CN110494752A

CN110494752A - With the method for early stage biomarker ubiquitin carboxy terminal hydrolase-l 1 assisted diagnosis measurement people experimenter traumatic brain injury degree

Info

Publication number: CN110494752A
Application number: CN201880020489.5A
Authority: CN
Inventors: B·麦奎斯顿; J·罗杰斯; S·德特维勒; J·马力诺
Original assignee: Abbott Laboratories
Current assignee: Abbott Laboratories
Priority date: 2017-03-23
Filing date: 2018-03-23
Publication date: 2019-11-22
Also published as: EP3602069A1

Abstract

Disclosed herein is use early stage biomarker ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) assisted diagnosis and evaluation suffer from or may suffer from damage to head, the human experimenter that such as slight or moderate is to severe trauma cerebral injury (TBI) method.It also discloses herein and the level based on UCH-L1 is helped to determine whether the subject that suffers from or may suffer from the damage to head will benefit from and therefore receive the method for head computerized tomography (CT) scanning.These methods are related to suffering from or may suffering from human experimenter the variation that the level and UCH-L1 level of UCH-L1 are detected in the one or more samples for being derived from the subject to the time point in after the damage on head 24 hours.

Description

With early stage biomarker ubiquitin carboxy terminal hydrolase-l 1 assisted diagnosis measurement people by The method of examination person's traumatic brain injury degree

Related application data

This application claims the U.S. Provisional Patent Application Serial No. 62/475,662 and 2018 submitted on March 23rd, 2017 The priority for the U.S. Patent Application Serial Number 15/934,541 that on March 23, in submits, content passes through for all purposes draws Be integrally incorporated herein.

Technical field

The present invention relates to by detecting when subject is suffered from or may be suffered from damage 24 hours to head Between point be derived from the early stage biomarker in one or more samples (such as one or more biological sample) of human experimenter The horizontal variation of object ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) carrys out assisted diagnosis and evaluation suffers from or may suffer from pair The damage on head, such as slight or moderate to severe trauma cerebral injury (TBI) human experimenter method.

Background technique

Only in the U.S., 5,000,000 mild trauma cerebral injuries (TBI) are just had more than every year and are occurred.Currently, without it is simple, Objective, accurate mensuration can be used for helping patient evaluation.In fact, many TBI evaluations and diagnosis are all based on subjective number According to.Unfortunately, such as the objective measurement approach of Cranial Computed Tomography and Glasgow coma score (GCS) is inadequate when evaluating slight TBI It is comprehensively or sensitive.In addition, Cranial Computed Tomography has no for most times of slight TBI, it is expensive, and make to suffer from Person is exposed to unnecessary radiation.In addition, negative Cranial Computed Tomography is not meant to that patient has been cleared by cerebral concussion；On the contrary, it can It is blue-sky for can be shown that certain intervening measures are such as performed the operation.It is objective that clinician and patient need, and reliable information is come accurate Such case is assessed, to promote appropriate point to examine and rehabilitation.So far, can be used for using UCH-L1 in acute care environment Help the data of evaluation of patient and management limited.

Slight TBI (or cerebral concussion) is more difficult to objectively detect and daily choose is presented in the critical care ward in the whole world War.Slight TBI will not usually cause general pathology, such as bleeding, and not have in the conventional computer tomoscan of brain It is abnormal, but the neuron dysfunction of rapid onset, subsided within a few days to a few weeks in spontaneous mode.About 15% Slight TBI patients' duration cognition dysfunction.For at the scene, in emergency ward and clinic, in sport area and There are outstanding demands for slight TBI victim in military activity (such as fight).

Summary of the invention

In one aspect, this disclosure relates to which a kind of assisted diagnosis and evaluation suffer from or may suffer from the damage to head The method of the human experimenter of (or head injury).The described method includes: a) being taken in 24 hours to after to head suspicious lesion It is measured from the biological sample of subject, to measure or detect ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in biological sample Level；And b) determine whether subject suffers from slight or moderate to severe trauma cerebral injury (TBI), wherein (i) working as life When UCH-L1 level in object sample is higher than the reference levels of UCH-L1, subject is determined as with moderate or severe trauma Cerebral injury, and when the UCH-L1 level in biological sample is lower than the reference levels of UCH-L1, subject is determined as suffering from Mild trauma cerebral injury；(ii) when from the UCH-L1 level in the biological sample that first time point obtains in the second time The UCH-L1 level in biological sample that point obtains when increasing or decreasing, subject is determined as suffering from statistically significant Have moderate or a severe trauma cerebral injury, and when from the UCH-L1 level in the biological sample that first time point obtains to Second time point obtain biological sample in UCH-L1 level it is not statistically significant when increasing or decreasing, by subject It is determined as with mild trauma cerebral injury；(iii) when the level of UCH-L1 subtracts from the first biological sample to the second biological sample When less or increasing at least absolute magnitude, subject is determined as with moderate or severe trauma cerebral injury, or when UCH-L1's When level does not decrease or increase at least absolute magnitude from the first biological sample to the second biological sample, subject is determined as suffering from Mild trauma cerebral injury；(iv) when the level of UCH-L1 decreases or increases at least from the first biological sample to the second biological sample When the first absolute magnitude, subject is determined as with moderate or severe trauma cerebral injury, or horizontal from the as UCH-L1 When one biological sample to the second biological sample decreases or increases at least the second absolute magnitude, subject is determined as with mild trauma Property cerebral injury；Or (v) when the UCH-L1 level in biological sample is higher than the reference levels of UCH-L1 or when from first Between to put UCH-L1 level in the biological sample of acquirement aobvious to the UCH-L1 level in the biological sample that the second time point obtained When work increases or decreases X amount, subject is determined as with moderate or severe trauma cerebral injury, and when in biological sample When UCH-L1 level is lower than the reference levels of UCH-L1 or as UCH-L1 from the biological sample that first time point obtains It, will when level is not dramatically increased or reduced greater than X amount to the UCH-L1 level in the biological sample that the second time point obtained Subject is determined as with mild trauma cerebral injury.

On the other hand, this disclosure relates to which a kind of assist in suffers from or may suffer from the doubtful damage to head Hurt the method for the traumatic brain injury degree in the human experimenter of (or head injury).The described method includes: a) to from tested At least two biological samples that person obtains are measured, and the first biological sample is derived from subject in 24 hours of suspicious lesion, And the second biological sample is derived from subject in about 3 to about 6 hours after obtaining the first biological sample；B) at least two biological samples The early stage biomarker of traumatic brain injury is detected in product, the early stage biomarker is by ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) it forms, wherein the presence of UCH-L1 starts to present in about 0 to about 6 hour after suspicious lesion；C) measurement first is raw Object sample and the second biological sample respectively in UCH-L1 it is horizontal, and determine that UCH-L1's is horizontal from the first biological sample to the Two biological samples are to reduce or increase；And d) level based on UCH-L1 is from the first biological sample to the second biological sample It reduces, increase and be also to maintain identical, determine the traumatic brain injury degree in subject.

In also yet other aspects, this disclosure relates to which a kind of assist in determining whether to suffering from or may suffer from doubtful The method that head computerized tomography (CT) scanning is carried out to the human experimenter of the damage (or head injury) on head.The method It include: that a) at least two biological samples obtained from subject are measured, the first biological sample is 24 small suspicious lesion When it is interior be derived from subject, and the second biological sample is derived from subject in about 3 to about 6 hours after obtaining the first biological sample；b) The early stage biomarker of traumatic brain injury is detected at least two biological samples, the early stage biomarker is by ubiquitin Carboxyl-terminal hydrolase L1 (UCH-L1) composition, wherein the presence of UCH-L1 start in about 0 to about 6 hour after suspicious lesion be in It is existing；C) measure the first biological sample and the second biological sample respectively in UCH-L1 it is horizontal, and determine UCH-L1 it is horizontal from First biological sample to the second biological sample is to reduce or increase；And it is d) horizontal from the first biological sample based on UCH-L1 To the second biological sample be reduce, increase also be to maintain it is identical, it is determined whether to subject carry out CT scan.

In yet other aspects, this disclosure relates to which a kind of assisted diagnosis and evaluation suffer from or may suffer from head injury Human experimenter method.For example, the method may include:

A) sample for being derived from subject in 24 hours after doubtful head injury is measured, to measure or detect sample The level of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in product；And

B) determine whether subject suffers from slight or moderate to severe trauma cerebral injury (TBI), in which:

(i) when the UCH-L1 level in sample is higher than the reference levels of UCH-L1, subject is determined as with moderate It is to severe trauma cerebral injury, and when the UCH-L1 level in sample is lower than the reference levels of UCH-L1, subject is true It is set to mild trauma cerebral injury；

(ii) when from the UCH-L1 level in the sample that first time point obtains to the sample obtained at the second time point In UCH-L1 level have statistically significant when increasing or decreasing, subject is determined as with moderate to severe trauma Property cerebral injury, and when from the UCH-L1 level in the sample that first time point obtains to the sample obtained at the second time point In UCH-L1 level it is not statistically significant when increasing or decreasing, by subject be determined as with mild trauma brain damage Wound；

It (iii), will be tested when the level of UCH-L1 decreases or increases at least absolute magnitude from the first sample to the second sample Person is determined as with moderate to severe trauma cerebral injury, or when the level of UCH-L1 does not have from the first sample to the second sample When decreasing or increasing at least absolute magnitude, subject is determined as with mild trauma cerebral injury；

It (iv), will be by when the level of UCH-L1 decreases or increases at least the first absolute magnitude from the first sample to the second sample Examination person is determined as with moderate to severe trauma cerebral injury, or when the level of UCH-L1 subtracts from the first sample to the second sample When less or increasing at least the second absolute magnitude, subject is determined as with mild trauma cerebral injury；Or

(v) when the UCH-L1 level in sample is higher than the reference levels of UCH-L1 or when obtaining from first time point Sample in UCH-L1 level dramatically increase or reduce X amount to the UCH-L1 level in the sample that the second time point obtained When, subject is determined as with moderate to severe trauma cerebral injury, and when the UCH-L1 level in sample is lower than UCH- It is obtained when the reference levels of L1 or when from the UCH-L1 level in the sample that first time point obtains at the second time point Sample in UCH-L1 level do not dramatically increase or reduce be greater than X amount when, by subject be determined as suffer from mild trauma Cerebral injury.

In yet other aspects, this disclosure relates to a kind of head injury (such as traumatic brain damage for evaluating human experimenter Wound) method.For example, the method may include:

A) sample (such as biological sample) for being derived from subject in 24 hours after doubtful head injury is measured, To measure the level of the ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in sample；And

(i) when the UCH-L1 level in sample is higher than the reference levels of UCH-L1, subject suffers from moderate to severe TBI, and wherein when the UCH-L1 level in sample is lower than the reference levels of UCH-L1, subject suffers from slight TBI；

(ii) when from the UCH-L1 level in the sample that first time point obtains to the sample obtained at the second time point In UCH-L1 level have statistically significant when increasing or decreasing, subject suffers from moderate to severe TBI, and its In when from the UCH-L1 level in the sample that first time point obtains to the UCH-L1 in the sample that the second time point obtained Horizontal not statistically significant when increasing or decreasing, subject suffers from slight TBI；

(iii) when the level of UCH-L1 decreases or increases at least absolute magnitude from the first sample to the second sample, subject Moderate is suffered to severe TBI, and wherein when the level of UCH-L1 is not decreased or increased from the first sample to the second sample At least absolute magnitude when, subject suffers from slight TBI；Or

(iv) tested when the level of UCH-L1 decreases or increases at least the first absolute magnitude from the first sample to the second sample Person suffers from moderate to severe TBI, and wherein when UCH-L1 it is horizontal from the first sample to the second sample decrease or increase to When few second absolute magnitude, subject suffers from slight TBI.

In the method for the head injury of any of above evaluation human experimenter, first time point can be doubtful head About 0 to about 12 hour after damage, and statistically significant increasing or decreasing can be selected from the group being made up of: (a) It is more than about 1 times from the second time point to first time point；(b) it is more than about 0.73 times from the second time point to first time point；Or (c) it is more than about 0.73 times from the second time point to first time point, and the reference levels of UCH-L1 are in about 350pg/mL and about Between 550pg/mL.In some embodiments, dramatically increase or reduce is from the second time point to first time point more than about 1 Times.In some embodiments, dramatically increase or reduce is from the second time point to first time point more than about 0.73 times.One In a little embodiments, dramatically increasing or reducing is to be more than about 0.73 times, and UCH-L1 from the second time point to first time point Reference levels between about 350pg/mL and about 550pg/mL.

In the method for any of above head injury for evaluating human experimenter, subject can be measured Before or after receive Golmud-Lhasa Pipeline scoring.For example, being scored based on Golmud-Lhasa Pipeline, subject may be doubted Like with moderate to severe TBI.Optionally, it is scored based on Golmud-Lhasa Pipeline, subject may be doubtful with slight TBI.Optionally, subject can not receive Golmud-Lhasa Pipeline scoring before being measured.In addition, subject It can receive Cranial Computed Tomography before being measured.Further optionally, subject can not receive before being measured Cranial Computed Tomography.Further, subject can not receive Golmud-Lhasa Pipeline or Cranial Computed Tomography before being measured. Further, subject can receive Golmud-Lhasa Pipeline before being measured but not receive Cranial Computed Tomography. Further, subject can receive Cranial Computed Tomography before being measured but not receive Golmud-Lhasa Pipeline. Further, subject can receive Cranial Computed Tomography and Golmud-Lhasa Pipeline before being measured.

In the method for any of above head injury for evaluating human experimenter, can by reference levels with suffer from The subject of moderate to severe TBI are associated (when compared with the subject with slight TBI).It is alternatively possible to will refer to It is horizontal associated (when compared with the subject for not suffering from TBI) with the subject with slight TBI.Further, it refers to Level can indicate subject with moderate to severe TBI (when compared with the subject with slight TBI).Further Ground, reference levels can indicate subject with slight TBI (when compared with the subject for not suffering from TBI).Further, It can be associated for 13-15 with Glasgow Glasgow coma scale by reference levels.Which kind of reference levels is selected to be used for the present invention It is apparent to a skilled reader in method.

In the method for any of above head injury for evaluating human experimenter, reference levels can (a) pass through Measuring method with the specificity between the sensibility and at least about 30% to about 100% between at least about 85% to about 100% It determines；(b) by having at least about 99% sensibility and the measuring method of at least about 75% specificity to determine；(c) at least About 50pg/mL is between about 12000pg/mL；Or (d) at least about 65pg/mL between about 9019pg/mL.In some implementations In mode, reference levels be can be by having the sensibility and at least about 30% between at least about 85% to about 100% to about What the measuring method of the specificity between 100% determined.In some embodiments, reference levels can be by having at least about 99% sensibility and at least about 75% specificity measuring method determine.In some embodiments, reference levels can be with In at least about 50pg/mL between about 12000pg/mL；Or (d) at least about 65pg/mL between about 9019pg/mL.

In the method for any of above head injury for evaluating human experimenter, sample can (a) in doubtful head It is obtained in about 0 to about 6 hour after portion's damage and reference levels is by the sensibility and at least about 33% with about 100% What the measuring method of specificity determined；(b) it is obtained in about 0 to about 6 hour after doubtful head injury and reference levels is to pass through What the measuring method with about 100% sensibility and about 100% specificity determined；(c) about 6 hours after doubtful head injury It is obtained between to about 12 hours and reference levels is the specificity by having about 100% sensibility and at least about 30% What measuring method determined；(d) it is obtained between after doubtful head injury about 6 hours to about 12 hours and reference levels is to pass through tool There is the measuring method of about 100% sensibility and at least about 63% specificity to determine；Or (e) about 6 after doubtful head injury It is obtained between hour to about 12 hours and reference levels is by the sensibility and at least about 96% at least about 90% What the measuring method of specificity determined.In some embodiments, sample can be after doubtful head injury in about 0 to about 6 hour It obtains and reference levels is determined by the measuring method with the specificity of about 100% sensibility and at least about 33%. In some embodiments, sample can obtain in about 0 to about 6 hour after doubtful head injury and reference levels are to pass through What the measuring method with about 100% sensibility and about 100% specificity determined.In some embodiments, sample can be with It is obtained between after doubtful head injury about 6 hours to about 12 hours and reference levels is by the sensitivity with about 100% Property and at least about 30% specificity measuring method determine.In some embodiments, sample can be in doubtful head injury It is obtained between about 6 hours to about 12 hours afterwards and reference levels is by the sensibility and at least about 63% with about 100% Specificity measuring method determine.In some embodiments, sample can be about 6 hours to about 12 after doubtful head injury It is obtained between hour and reference levels is the survey by having the specificity of at least about 90% sensibility and at least about 96% Determine what method determined.

Optionally, sample can (a) after doubtful head injury in 0 to about 6 hour obtain and reference levels be about 311pg/mL；(b) it is obtained in 0 to 6 hour after doubtful head injury and reference levels is about 9019pg/mL；(c) doubtful It is obtained between about 6 hours to about 12 hours after head injury and reference levels is about 98pg/mL；(d) in doubtful head injury It is obtained between about 6 hours to about 12 hours afterwards and reference levels is about 209pg/mL；Or (e) about 6 after doubtful head injury It is obtained between hour to about 12 hours and reference levels is about 569pg/mL.In some embodiments, sample can doubt Like obtaining in after head injury 0 to about 6 hour and reference levels are about 311pg/mL.In some embodiments, sample can With after doubtful head injury in 0 to 6 hour obtain and reference levels be about 9019pg/mL.In some embodiments, sample Product can between after doubtful head injury about 6 hours to about 12 hours it is interior acquirement and reference levels be about 98pg/mL.One In a little embodiments, sample can between after doubtful head injury about 6 hours to about 12 hours interior acquirement and reference levels It is about 209pg/m.In some embodiments, sample can be interior between after doubtful head injury about 6 hours to about 12 hours It obtains and reference levels is about 569pg/mL.

In the method for any of above head injury for evaluating human experimenter, can by reference levels with suffer from The subject of moderate to severe TBI are associated (when compared with the subject with slight TBI).It is alternatively possible to will refer to It is horizontal associated (when compared with the subject for not suffering from TBI) with the subject with slight TBI.Further, it refers to Level can indicate subject with moderate to severe TBI (when compared with the subject with slight TBI).Further Ground, reference levels can indicate subject with slight TBI (when compared with the subject for not suffering from TBI).Still further Ground, can be associated for 13-15 with Glasgow Glasgow coma scale by reference levels.Select which kind of reference levels for this hair It is apparent to a skilled reader in bright method.

In the method for any of above head injury for evaluating human experimenter, absolute magnitude be can be by having The measuring method of specificity between sensibility and at least about 30% to about 100% between at least about 70% to about 100% determines 's.

In the method for any of above head injury for evaluating human experimenter, sample can (a) in doubtful head Portion damage after in about 0 to about 6 hour obtain and absolute magnitude be by with about 100% sensibility and about 100% it is special Property measuring method determine；(b) it is obtained in about 6 to about 12 hours after doubtful head injury and absolute magnitude is by having extremely Lack the measuring method determination of about 70% sensibility and at least about 92% specificity；(c) after doubtful head injury about 0 to about It is obtained in 10 hours and absolute magnitude is true by the measuring method with the specificity of about 100% sensibility and at least about 36% Fixed；(d) it is obtained in about 0 to about 11 hour after doubtful head injury and absolute magnitude is by the sensitivity with about 100% Property and at least about 32% specificity measuring method determine；Or it (e) is obtained in about 0 to about 12 hour after doubtful head injury And absolute magnitude is determined by the measuring method with the specificity of at least about 75% sensibility and at least about 76%.

Further, in the method for any of above head injury for evaluating human experimenter, sample can be with (a) it is obtained in about 0 to about 6 hour after doubtful head injury and absolute magnitude is about 2528pg/mL；(b) it is damaged on doubtful head It is obtained in about 6 to about 12 hours after wound and absolute magnitude is about 129pg/mL；(c) about 6 to about 12 small after doubtful head injury When it is interior acquirement and absolute magnitude be about 25pg/mL；(d) it is obtained in 0 to 11 hour after doubtful head injury and absolute magnitude is About 25pg/mL；Or (e) after doubtful head injury in about 0 to about 12 hour obtain and absolute magnitude be about 129pg/mL.

In the method for the head injury for evaluating human experimenter confirmed above, sample (or biological sample) is complete Blood sample, blood serum sample or plasma sample.Specifically, sample (or biological sample) can be whole blood sample.Optionally, sample (or biological sample) can be blood serum sample.Optionally, sample (or biological sample) can be plasma sample.

In the method for the above-mentioned head injury for evaluating human experimenter, UCH-L1 is surveyed using immunoassay It measures or detects.In one aspect, in the above-mentioned methods, sample (or biological sample) can be whole blood sample and UCH-L1 is Use Immunoassays measure or detection.In yet other aspects, in the above-mentioned methods, sample (or biological sample) be can be Blood serum sample and UCH-L1 are using Immunoassays measure or detection.In still another aspect, in the above-mentioned methods, sample Product (or biological sample) can be plasma sample and UCH-L1 is using Immunoassays measure or detection.

The method of the above-mentioned head injury for evaluating human experimenter may further include with the therapy for being directed to TBI Treatment is assessed as the human experimenter with slight or moderate to severe TBI.The such a patient for TBI treatment may be used also To be optionally monitored during and after any course for the treatment of.Optionally, it is evaluated to may further include monitoring for the method For with slight or moderate TBI human experimenter (may such as not receive those of any treatment yet so far).

In yet other aspects, this disclosure relates to which a kind of assist in the mankind for suffering from or may suffering from head injury The method of traumatic brain injury degree in subject.For example, the method may include:

A) at least two samples obtained from subject are measured, the first sample is 24 small doubtful head injury When it is interior be derived from subject, and the second sample is derived from subject in about 3 to about 6 hours after obtaining the first sample；

B) the early stage biomarker of traumatic brain injury, the early stage biomarker are detected at least two samples It is made of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1)；

C) measure the first sample and the second sample respectively in UCH-L1 it is horizontal, and determine that UCH-L1's is horizontal from first Sample is to reduce or increase to the second sample；And

D) based on UCH-L1 it is horizontal from the first sample to the second sample be reduce, increase also be to maintain it is identical, determine by Traumatic brain injury degree in examination person, wherein determining that traumatic brain injury degree includes determining whether subject suffers from slightly Or moderate is to severe trauma cerebral injury.

In above for the method for assisting in the traumatic brain injury degree in human experimenter, UCH-L1 is (such as Presence in stepb) starts to present after doubtful head injury in about 0 to about 6 hour.

In yet other aspects, this disclosure relates to traumatic brain injury (TBI) degree in a kind of evaluation human experimenter Method.For example, the method may include:

A) it is measured with the quantitative ubiquitin carboxy terminal hydrolase-l 1 from least two samples that subject obtains (UCH-L1), the first sample is derived from subject in 24 hours of doubtful head injury, and the second sample is obtaining the first sample It is derived from subject within about 3 to about 6 hours after product；With

B) based on UCH-L1 it is horizontal from the first sample to the second sample be reduce, increase also be to maintain it is identical, determine by TBI degree in examination person, wherein determining that traumatic brain injury degree includes determining whether subject suffers from slight or moderate extremely Severe trauma cerebral injury.

In the above-mentioned method for determining or evaluating TBI degree, the first sample can be about 0 after doubtful head injury It is obtained in about 6 hours.

It is horizontal from the first sample to second as UCH-L1 in the above-mentioned method for determining or evaluating TBI degree When sample increases or decreases at least about 20pg/mL at least about 6100pg/mL, subject can be determined with moderate to severe TBI。

In the above-mentioned method for determining or evaluating TBI degree, the first sample can (a) after doubtful head injury It is obtained in about 0 to about 6 hour and the level of UCH-L1 increases or decreases at least about 2528pg/mL；(b) in doubtful head injury It is obtained in about 6 to about 12 hours afterwards and the level of UCH-L1 increases or decreases at least about 129pg/mL；(c) it is damaged on doubtful head It is obtained in about 0 to about 10 hour after wound and the level of UCH-L1 increases or decreases at least about 25pg/mL；(d) on doubtful head It is obtained in about 0 to about 11 hour after damage and the level of UCH-L1 increases or decreases at least about 25pg/mL；Or (e) doubtful It is obtained in about 0 to about 12 hour after head injury and the level of UCH-L1 increases or decreases at least about 129pg/mL.

In the method for determining or evaluating TBI degree confirmed above, sample (or biological sample) can be whole blood Sample, blood serum sample or plasma sample.Specifically, sample (or biological sample) can be whole blood sample.Optionally, sample (or Biological sample) it can be blood serum sample.Optionally, sample (or biological sample) can be plasma sample.

In the above-mentioned method for determining or evaluating TBI degree, UCH-L1 is using Immunoassays measure or inspection It surveys.On the other hand, in the above-mentioned methods, sample (or biological sample) can be whole blood sample and UCH-L1 be using Immunoassays measure or detection.On the other hand, in the above-mentioned methods, sample (or biological sample) can be serum sample Product and UCH-L1 are using Immunoassays measure or detection.On the other hand, in the above-mentioned methods, sample is (or raw Object sample) it can be plasma sample and UCH-L1 is using Immunoassays measure or detection.

The above-mentioned method for determining or evaluating TBI degree may further include with the therapy treatment quilt for TBI It is evaluated as the human experimenter with slight or moderate to severe TBI.The such a patient for TBI treatment can also be optional Ground is monitored as described during and after any course for the treatment of.Optionally, the method may further include prison It surveys and is assessed as (may such as not receiving that of any treatment yet so far with slight or moderate TBI human experimenter A bit).

In yet other aspects, this disclosure relates to which one kind assists in determining whether to suffering from or may suffer from head injury Human experimenter carry out head computerized tomography (CT) scanning method.For example, the method may include: be measured with Detection measures from the ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) at least two samples that subject obtains, the first sample Product are derived from subject in 24 hours of doubtful head injury, and the second sample is about 3 to about 6 hours after obtaining the first sample It is derived from subject；

A) measure the first sample and the second sample respectively in UCH-L1 it is horizontal, and determine that UCH-L1's is horizontal from first Sample is to reduce or increase to the second sample；And

B) horizontal based on UCH-L1 is to reduce, increase and be also to maintain identical from the first sample to the second sample, and determination is It is no that CT scan is carried out to subject, in which:

(i) CT scan is carried out when the UCH-L1 level in the first sample is higher than the reference levels of UCH-L1, and wherein When the UCH-L1 level in the first sample is lower than the reference levels of UCH-L1 without CT scan；

(ii) statistically aobvious when having from the UCH-L1 level in the first sample to the UCH-L1 level in the second sample Write CT scan is carried out when increasing or decreasing, and wherein when from the UCH-L1 level in the first sample into the second sample UCH-L1 level it is not statistically significant when increasing or decreasing without CT scan；Or

(iii) CT is carried out when the level of UCH-L1 decreases or increases at least absolute magnitude from the first sample to the second sample to sweep Retouch, and wherein when UCH-L1 level do not decrease or increase at least absolute magnitude from the first sample to the second sample when without CT scan.

In yet other aspects, this disclosure relates to which one kind evaluates whether to carry out head computerized tomography to human experimenter (CT) method scanned.For example, the method may include:

A) it is measured with quantitative from the ubiquitin carboxy terminal hydrolase-l 1 at least two samples that subject obtains (UCH-L1), the first sample is derived from subject in about 24 hours of doubtful head injury, and the second sample is obtaining first It is derived from subject within about 3 to about 6 hours after sample；With

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, first time point can be doubtful head About 0 to about 12 hour after damage, and statistically significant increasing or decreasing is from first time point to the second time point Less than about 2 times；First time point can be about 0 to about 12 hour after doubtful head injury, and statistically significant increasing Add deduct is to be less than about 1.81 times from first time point to the second time point less；After first time point can be doubtful head injury About 0 to about 12 hour, and statistically significant increase is from the second time point to first time point more than about 0.50 times； First time point can be about 0 to about 12 hour after doubtful head injury, and statistically significant increase is from second Time point is more than about 0.55 times to first time point；Or first time point can be about 0 to about 12 after doubtful head injury Hour, the reference levels of UCH-L1 are about 550pg/mL, and statistically significant increase is from the second time point to first Time point is more than about 0.55 times.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, based on the CT scan carried out, by Examination person doubtful may suffer from traumatic brain injury.For example, according to the medical conditions (such as if patient unconscious) of subject, CT scan can be carried out soon to assess and/or evaluate subject after subject reaches emergency ward, trauma center or other places Whether TBI is suffered from.Can carry out for confirm and determine subject whether suffer from slight or moderate or severe TBI measurement it It is preceding to carry out this CT scan.After being measured, one or many subsequent CT scan can be carried out based on measurement result, with The a part managed as doctor (or other medical workers) TBI is (such as to determine whether that operation and/or medicine may be needed Object intervention).

It, can be by reference levels and male portion CT in the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography Scanning is associated.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, reference levels can (a) by having The measuring method of specificity between sensibility and at least about 30% to about 100% between at least about 80% to about 100% determines； (b) at least about 50pg/mL between about 1000pg/mL；Or (c) at least about 86pg/mL between about 700pg/mL.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, sample can (a) damaged on doubtful head It is obtained in about 0 to about 6 hour after wound and reference levels is by having about 100% sensibility and at least about 37.5% spy What anisotropic measuring method determined；(b) it is obtained in about 0 to about 6 hour after doubtful head injury and reference levels is to pass through tool There is the measuring method of about 100% sensibility and at least about 75% specificity to determine；(c) about 6 small after doubtful head injury It is obtained between about 12 hours and reference levels is by having at least about 96% sensibility and at least about 30% spy What anisotropic measuring method determined；(d) it is obtained between after doubtful head injury about 6 hours to about 12 hours and reference levels is By having the measuring method of at least about 86% sensibility and at least about 35% specificity to determine；Or (e) on doubtful head It is obtained between about 0 hour to about 12 hours after damage and reference levels is by the sensibility and at least about with about 100% What the measuring method of 32% specificity determined.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, sample can (a) damaged on doubtful head It is obtained in about 0 to about 6 hour after wound and reference levels is about 370pg/mL；(b) about 0 to about 6 small after doubtful head injury When it is interior acquirement and reference levels be about 509pg/mL；(c) it is obtained between after doubtful head injury about 6 hours to about 12 hours And reference levels are about 96pg/mL；(d) it obtains and refers between after doubtful head injury about 6 hours to about 12 hours Level is about 86pg/mL；Or (e) between after doubtful head injury about 6 hours to about 12 hours obtain and reference levels be About 550pg/mL.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, by absolute magnitude and male portion CT scan phase Association.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, absolute magnitude be can be by having at least What the measuring method of the specificity between the sensibility and at least about 30% to about 100% between about 80% to about 100% determined.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, sample can be with: (a) damaging on doubtful head It is obtained in about 0 to about 10 hour after wound and absolute magnitude is by having at least about 85% sensibility and at least about 41% spy What anisotropic measuring method determined；Or (b) after doubtful head injury in about 0 to about 10 hour obtain and absolute magnitude be to pass through tool There is the measuring method of at least about 90% sensibility and at least about 35% specificity to determine.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, sample can (a) damaged on doubtful head It is obtained in about 0 to about 10 hour after wound and absolute magnitude is about 25pg/mL；Or it is (b) about 0 to about 10 small after doubtful head injury When it is interior acquirement and absolute magnitude be about 23pg/mL.

Above-mentioned uses the therapy for being directed to TBI for determining or evaluating whether that the method for carrying out Cranial Computed Tomography may further include Treatment is assessed as the human experimenter with slight or moderate to severe TBI.The such a patient for TBI treatment may be used also To be optionally monitored during and after any course for the treatment of.Optionally, it is evaluated to may further include monitoring for the method For with slight or moderate TBI human experimenter (may such as not receive those of any treatment yet so far).

In the method for determining or evaluating whether to carry out Cranial Computed Tomography confirmed above, sample (or biological sample) is complete Blood sample, blood serum sample or plasma sample.Specifically, sample (or biological sample) can be whole blood sample.Optionally, sample (or biological sample) can be blood serum sample.Optionally, sample (or biological sample) can be plasma sample.

In the above-mentioned method for determining or evaluating whether to carry out Cranial Computed Tomography, UCH-L1 be can be using immunoassays What method was measured or was detected.In the other side of the above method, sample (or biological sample) can be whole blood sample and UCH- L1 can be using Immunoassays measure or detection.Other side in the above-mentioned methods, sample (or biological sample) It can be blood serum sample and UCH-L1 can be using Immunoassays measure or detection.In the above-mentioned methods another Aspect, sample (or biological sample) can be plasma sample and UCH-L1 can be using Immunoassays measure or detection 's.

In any above method, in some embodiments, the second sample after doubtful head injury about 5 hours to about Subject is obtained between 16 hours or about 3 hours to about 6 hours.More specifically, the second sample can be after doubtful head injury About 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours or about It is derived from subject within 12 hours.

Any of above method, which may further include, to be measured sample to measure or detect the one kind for not being UCH-L1 Or the level of a variety of other biomarkers.

In any above method, measurement may include:

(a) level of one of the first sample and the second sample or a variety of other biomarkers is measured or detects,

(b) determine the level of one or more other biomarkers from the first sample to the reduction of the second sample or increasing Add, and

If (c) one of from one of first sample or a variety of other biomarker levels to the second sample or A variety of other biomarker levels are decreased or increased with statistically significant, then subject are evaluated as with slight TBI。

In the above-mentioned methods, one or more other biomarkers are selected from the group being made up of: S100 β, neuron Specificity olefinic alcohol enzyme (NSE), glial fibrillary acidic protein (GFAP), Apo lipoprotein 1, Tau, C reactive protein (CRP), Free brain-derived neurotrophic factor (BDNF), p-Tau, total BDNF, Troponin I (TnI) and combinations thereof.More specifically, one Kind or a variety of other biomarkers are S100 β, BDNF or CRP.

In any above method, the level of UCH-L1 be can be by carrying out immunoassays measurement.For example, UCH-L1 Level can be and in the following manner measure:

(a) make sample simultaneously or successively contacted in any order with following:

(1) antibody is captured, in conjunction with the epitope in UCH-L1 or UCH-L1 segment to form capture antibody-UCH-L1 antigen Compound, and

(2) antibody is detected, it includes detectable label and combines the epitope at large for obtaining antibody combination on UCH-L1, with UCH-L1 antigen-detection antibody complex is formed,

So that capture antibody-UCH-L1 antigen-detection antibody complex is formed, and

(b) based on the letter generated by the detectable label in capture antibody-UCH-L1 antigen-detection antibody complex Number, measure the amount or concentration of UCH-L1 in sample.

In any above method, sample can be whole blood sample, blood serum sample, spinal fluid samples or plasma sample.In On one side, sample is whole blood sample.On the other hand, sample is blood serum sample.In yet other aspects, sample is blood plasma Sample.This sample can obtain in many ways.For example, can subject by shaken by body, cause closed or Passivity impact, one or many tumbles, explosion or the shock wave that the external mechanical force of open head trauma or other power generate Or sample is obtained after head injury caused by other types of blunt forces wound.It is alternatively possible to take in or expose in subject Sample is obtained after the combination of chemical substance, toxin or chemical substance and toxin.Chemical substance or the example of toxin are fiery, mould Bacterium, asbestos, insecticide, insecticide, organic solvent, paint, glue, gas, organic metal, drug abuse or its one kind or more Kind combination.Still further, sample can be from suffering from autoimmune disease, metabolic disorder, brain tumor, anoxic, virus, meninx The subject of scorching, hydrocephalus or combinations thereof obtains.

Any of above method can carry out on anyone class subject, without considering in the group being made up of Factor: slight or moderate is suffered from the conduct of the clinical condition of human experimenter, the laboratory evaluation of human experimenter, human experimenter Head may be suffered to the classification of severe TBI, the performance of the low or high UCH-L1 level of human experimenter and human experimenter The timing of any event of portion's damage.

Detailed description of the invention

Fig. 1 shows the biomarker UCH-L1 result relative to the time from damage.

Fig. 2 shows according to recipient's operating characteristics (ROC) to UCH-L1 level associated with CT at time point point Analysis.

Fig. 3, which is shown, changes (" increment " compared to the UCH-L1 level at time point 1 to time point 2 associated with CT (i.e. time point 2- time point 1)) recipient's operating characteristics (ROC) analysis.

Fig. 4 is shown to the absolute magnitude (" absolute increment ") of UCH-L1 result associated with CT (i.e. at time point 2 The absolute difference of UCH-L1 level at UCH-L1 level and time point 1) recipient's operating characteristics (ROC) analysis.

Fig. 5 shows the box-shaped figure of the UCH-L1 measurement result according to time point.

Fig. 6 shows the measurement dynamics intermediate value Z-score according to the test result at time point.

Fig. 7 A show with it is positive relative to negative CT scan result it is associated time point 1 (after head injury 0 to Obtained in 6 hours) and time point 2 (after 1 sample of time point 3 to 6 hours obtain) UCH-L1 measurement result box-shaped figure.Figure 7B is shown and the positive UCH-L1 measurement result relative at negative CT scan result associated time point 1 and time point 2 ROC curve.

Fig. 8 A show with it is positive relative to negative CT scan result it is associated time point 1 (after head injury 6 to Obtained in 12 hours) and time point 2 (after 1 sample of time point 3 to 6 hours obtain) UCH-L1 measurement result box-shaped figure. Fig. 8 B is shown to be tied with the positive UCH-L1 measurement relative at negative CT scan result associated time point 1 and time point 2 The ROC curve of fruit.

Fig. 9 A and Fig. 9 B are shown according to the time range for obtaining the first sample with the positive relative to negative CT scan knot The box-shaped figure of difference or variation (" increment ") UCH-L1 measurement result between fruit associated time point 2 and time point 1.Fig. 9 A The box-shaped figure and Fig. 9 B shown for 0-6 hours groups shows the box-shaped figure organized for 6-12 hours.

Figure 10 A and Figure 10 B are shown according to the time range for obtaining the first sample with the positive relative to negative CT scan As a result the absolute magnitude (" absolute increment ") (absolute difference i.e. between time point 2 and time point 1) of associated UCH-L1 result Box-shaped figure.Figure 10 A shows the box-shaped figure organized for 0-6 hours and Figure 10 B shows the box-shaped figure for 6-12 hours groups.

Figure 11 A show with slightly relative to moderate/severe TBI GCS scoring associated time point 1 (in head injury Obtained in 0 to 6 hour afterwards) and time point 2 (after 1 sample of time point 3 to 6 hours obtain) at UCH-L1 measurement result case Shape figure.Figure 11 B shows and slightly scores at associated time point 1 and time point 2 relative to moderate/severe TBI GCS The ROC curve of UCH-L1 measurement result.

Figure 12 A show with slightly relative to moderate/severe TBI GCS scoring it is associated time point 1 (head damage Obtained in 6 to 12 hours after wound) and time point 2 (after 1 sample of time point 3 to 6 hours obtain) UCH-L1 measurement result Box-shaped figure.Figure 12 B show with slightly relative to moderate/severe TBI GCS scoring associated time point 1 and time point 2 at UCH-L1 measurement result ROC curve.

Figure 13 A and Figure 13 B show according to obtain the first sample time range with slightly relative to moderate/severe TBI GCS scores the difference between associated time point 2 and time point 1UCH-L1 measurement result or the case of variation (" increment ") Shape figure.Figure 13 A shows the box-shaped figure organized for 0-6 hours and Figure 13 B shows the box-shaped figure for 6-12 hours groups.

Figure 14 A show with it is slightly absolute relative to the associated UCH-L1 measurement result of moderate/severe TBI GCS scoring Measuring (" absolute increment "), (i.e. time point 1 (obtaining in 0 to 6 hour after head injury) and time point 2 is (after 1 sample of time point 3 to 6 hours obtain) between absolute difference) box-shaped figure.Figure 14 B shows and slightly comments relative to moderate/severe TBI GCS The ROC curve of the associated UCH-L1 measurement result absolute magnitude of split-phase (" absolute increment ").

Figure 15 A shows and slightly scores the absolute magnitude of associated UCH-L1 result relative to moderate/severe TBI GCS (" absolute increment ") (i.e. time point 1 (obtaining in 6 to 12 hours after head injury) and time point 2 is (3 after 1 sample of time point To 6 hours obtain) between absolute difference) box-shaped figure.Figure 15 B shows and slightly scores relative to moderate/severe TBI GCS The ROC curve of the absolute magnitude (" absolute increment ") of associated UCH-L1 result.

Figure 16 show with slightly relative to moderate/severe TBI GCS scoring associated time point 1 (in head injury Obtained in 0 to 6 hour afterwards) and time point 2 (after 1 sample of time point 3 to 6 hours obtain) between UCH-L1 level multiple The ROC curve of variation.

Figure 17 shows the absolute magnitude of UCH-L1 result associated with CT scan (" absolute increment ") (i.e. 1 (In of time point Obtained in 0 to 10 hour after head injury) and time point 2 (after 1 sample of time point 3 to 6 hours obtain) between absolute difference) ROC curve.

Figure 18 shows absolute magnitude (" absolute increment ") (i.e. 1 (In of time point of UCH-L1 result associated with CT scan After head injury more than 10 hours obtain) with time point 2 (after 1 sample of time point 3 to 6 hours acquirement) between absolute difference) ROC curve.

Figure 19 shows and slightly scores the absolute magnitude of associated UCH-L1 result relative to moderate/severe TBI GCS (" absolute increment ") (i.e. time point 1 (obtaining in 0 to 11 hour after head injury) and time point 2 is (3 after 1 sample of time point To 6 hours obtain) between absolute difference) ROC curve.

Figure 20 shows and slightly scores the absolute magnitude of associated UCH-L1 result relative to moderate/severe TBI GCS (" absolute increment ") (i.e. time point 1 (obtaining after head injury more than 11 hours) and time point 2 is (3 after 1 sample of time point To 6 hours obtain) between absolute difference) ROC curve.

Figure 21 shows and slightly scores the absolute magnitude of associated UCH-L1 result relative to moderate/severe TBI GCS (" absolute increment ") (i.e. time point 1 (obtaining in 0 to 10 hour after head injury) and time point 2 is (3 after 1 sample of time point To 6 hours obtain) between absolute difference) ROC curve.

Figure 22 shows and slightly scores the absolute magnitude of associated UCH-L1 result relative to moderate/severe TBI GCS (" absolute increment ") (i.e. time point 1 (obtaining after head injury more than 12 hours) and time point 2 is (3 after 1 sample of time point To 6 hours obtain) between absolute difference) ROC curve.

Specific embodiment

The present invention relates to use early stage biomarker ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) assisted diagnosis and comment Valence suffers from the damage to head, subject (the such as mankind of such as slight or moderate to severe trauma cerebral injury (TBI) Subject) method.When these methods are related to detecting the difference in 24 hours to head injury or to head suspicious lesion Between to put the UCH-L1 that is derived from one or more samples (such as biological sample) of subject horizontal.To head injury or doubt Like detecting the increase of UCH-L1 level in after damage initial 24 hours or increasing, the subsequent reduction of UCH-L1 level can be later So that accurate evaluation or diagnosis subject are helped, to allow subsequent early treatment and monitoring with slight or moderate to severe The patient of traumatic brain injury.For example, following subjects can be accredited as with moderate to severe trauma cerebral injury: described Subject has the UCH- in the first sample in initial 6 hours, obtained in such as 2 to 6 hours after damage or suspicious lesion L1 level is compared to about 0.5 to 10 hour after obtaining the first sample, the statistics of such as 3 to 6 hours the second samples obtained Upper significant change.In another example, the subject with the UCH-L1 level for being higher than reference levels can also be identified For with moderate to severe trauma cerebral injury.In another example, the increasing at least with absolute magnitude can also be added deduct Few subject is accredited as with moderate to severe trauma cerebral injury.

The invention further relates to helping the level based on UCH-L1 to determine, whether the subject for suffering from the damage to head will The method for benefiting from and therefore receiving head computerized tomography (CT) scanning.These methods are related to detection to head injury or right Different time points in 24 hours of head suspicious lesion are derived from the level of the UCH-L1 in one or more samples of subject.In To detecting the increase of UCH-L1 level in after head injury or suspicious lesion initial 24 hours or increasing, UCH-L1 can be later Horizontal subsequent reduction is so that assist in whether subject should receive head CT scan.For example, following subjects can be reflected Positive Cranial Computed Tomography may be had and therefore benefit from carry out CT scan by being set to: the subject has in damage or suspicious lesion Afterwards in initial 6 hours, the UCH-L1 level in the first sample obtained in such as 2 to 6 hours is compared in the first sample of acquirement About 0 to 10 hour afterwards, the statistically significant variation of such as 3 to 6 hours the second samples obtained.In another example, Can also will have higher than reference levels UCH-L1 level subject be accredited as may have positive head CT scan and Therefore carry out CT scan is benefited from.In another example, at least can also there will be increasing or decreasing for absolute magnitude tested Person is accredited as to have positive head CT scan (for example, thereby indicate that potential TBI) and therefore benefit from and carries out CT and sweep It retouches.

Chapter title used in this section and the entire disclosure of this paper is only used for organizational goal and is not intended to limit Property.

1、Definition

Unless otherwise defined, otherwise all technical and scientific terms used herein has and those of ordinary skill in the art The identical meaning being generally understood.When the conflict occurs, it is subject to this document (including definition).Although in practice of the invention Or the method and material similar or equivalent with method those of described herein and material can be used in test, but be described below Preferred method and material.Side of all announcements, patent application, patent and other bibliography being mentioned above to quote Formula is integrally incorporated herein.Material, method and embodiment disclosed herein are merely exemplary and are not intended to restrictive.

As used herein, term "comprising", " comprising ", " having ", " having ", " can with ", " containing " and its version Open conjunction, term or the words for a possibility that being intended to be not excluded for other behavior or structure.Unless context is in addition clear Ground regulation, otherwise singular "one", "an" and " described " include plural reference.The disclosure also covers " including this paper institute The embodiment or element of presentation ", " being made of embodiment presented herein or element " and " mainly by presented herein Embodiment or element composition " other embodiment, regardless of whether being explicitly described.

For the narration of this paper numberical range, clearly cover with same accuracy it is intervenient it is each in Between number.For example, also covering number 7 and 8 in addition to 6 and 9 for range 6-9, and for range 6.0-7.0, clearly cover Number 6.0,6.1,6.2,6.3,6.4,6.5,6.6,6.7,6.8,6.9 and 7.0.

" affinity maturation antibody " has the anti-of one or more variations for referring to herein in one or more CDR Body, the variation cause the antibody compared with the parental antibody for not having the variation for affinity (the i.e. K of target antigen_D、 k_dOr k_a) improve.The parent that exemplary affinity maturation antibody will have target antigen nanomolar concentration or even picomolar concentrations And power.Multiple programs for generating affinity maturation antibody are well known in the art, including to the biological display system of use The screening of standby combinatorial antibody library.For example, Marks et al., BioTechnology 10:779-783 (1992) describe it is logical Cross the affinity maturation that the reorganization of VH and VL structural domain carries out.To the random mutagenesis of CDR and/or Framework residues by being described below: Barbas et al., Proc.Nat.Acad.Sci.USA, 91:3809-3813 (1994)；Schier et al., Gene, 169:147- 155(1995)；Yelton et al., J.Immunol., 155:1994-2004 (1995)；Jackson et al., J.Immunol., 154(7):3310-3319(1995)；With Hawkins et al., J.Mol.Biol., 226:889-896 (1992).It is lured selectively Displacement sets and is described in United States Patent (USP) by the selective mutation that increased activity amino acid residue carries out in contact or super mutated site Number 6,914,128 B1.

As used herein " a kind of antibody " and " Multiple Antibodies " refer to monoclonal antibody, multi-specificity antibody, human antibody, Humanized antibody (complete or partial humanization), animal's antibody such as, but not limited to bird (such as duck or goose), shark, whale and Mammal (including non-primate (such as ox, pig, camel, yamma, horse, goat, rabbit, sheep, hamster, cavy, Cat, dog, rat, mouse etc.) or non-human primate (such as monkey, chimpanzee etc.)), it is recombinant antibodies, chimeric antibody, single-stranded Fv (" scFv "), single-chain antibody, single domain antibody, Fab segment, F (ab') segment, F (ab')₂Segment, disulfide bond connect Fv (" sdFv ") and antiidiotype (" anti-Id ") antibody, bi-domain antibody, double variable domains (DVD) or three variable domains (TVD) (double variable domains immunoglobulins and preparation method thereof are described in Wu, C et al., Nature to antibody Biotechnology, 25 (11): 1290-1297 (2007) and PCT international application WO 2001/058956, each document it is interior Hold be incorporated herein by reference) and any of above antibody functional activity epitope binding fragments.Specifically, antibody includes The immunoreactive fragments of immunoglobulin molecules and immunoglobulin molecules, the i.e. molecule containing analyte binding site.It is immune Globulin molecule can have any type (such as IgG, IgE, IgM, IgD, IgA and IgY), classification (such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass.It for the sake of simplicity, is herein commonly referred to as " anti-point for the antibody of analyte Analyse object antibody " or be only " analyte antibody " (such as anti-UCH-L1 antibody or UCH-L1 antibody).

" antibody fragment " refers to a part of the complete antibody comprising antigen binding site or variable region as used herein. The part does not include the constant heavy structural domain (i.e. CH2, CH3 or CH4 depend on antibody isotype) in the area complete antibody Fc. The example of antibody fragment includes but is not limited to Fab segment, Fab' segment, Fab'-SH segment, F (ab')₂Segment, Fd segment, Fv Segment, scFv (scFv) molecule, the single chain polypeptide only containing a light variable domains, contains light chain variable at double antibody The single chain polypeptide of three CDR of structural domain, the only single chain polypeptide containing heavy chain variable region and contain heavy chain variable region The single chain polypeptide of three CDR.

" area under the curve " or " AUC " refers to the area under ROC curve.AUC under ROC curve is the measurement of accuracy. AUC is the perfect test of 1 expression, and AUC indicates meaningless test for 0.5.Preferred AUC can be at least about 0.700, extremely Few about 0.750, at least about 0.800, at least about 0.850, at least about 0.900, at least about 0.910, at least about 0.920, at least about 0.930, at least about 0.940, at least about 0.950, at least about 0.960, at least about 0.970, At least about 0.980, at least about 0.990 or at least about 0.995.

" bead " and " particle " is used interchangeably herein, and refers to made of substantially spherical solid support.Bead Or an example of particle is particle.The particle that can be used for this paper can be any type as known in the art.For example, bead Or particle can be magnetic bead or magnetic-particle.Magnetic beads/particle can be ferromagnetic, ferrimagnetism, paramagnetic, super It is paramagnetic or ferrofluid.Exemplary ferromagnetic material includes Fe, Co, Ni, Gd, Dy, CrO₂, MnAs, MnBi, EuO and NiO/Fe.The example of ferrimagnetic material includes NiFe₂O₄、CoFe₂O₄、Fe₃O₄(or FeO'Fe₂O₃).Bead can have magnetic It solid core part and is surrounded by one or more nonmagnetic layers.Optionally, magnetic part can be around non-magnetic core Layer.Particle can have any size to work in methods described herein, and for example, about 0.75 to about 5nm or about 1 to about 5nm or about 1 to about 3nm.

" binding protein " is herein for referring to monomer or poly with binding partner binds and formed compound Body protein matter, such as polypeptide, antigen, chemical compound or other molecules or any kind of substrate.Binding protein specificity In conjunction with binding partners.Binding protein includes antibody and its antigen-binding fragment and its as known in the art and hereafter institute The various other forms and derivative stated, and include one or more specific sites combined on antigen molecule or antigen molecule Other molecules of the antigen-binding domains of (epitope).Therefore, binding protein includes but is not limited to antibody, tetramer immune globulin White, IgG molecule, IgG1 molecule, monoclonal antibody, chimeric antibody, CDR grafted antibody, humanized antibody, affinity maturation are anti- The reservation of body and any such antibody combines the segment of the ability of antigen.

" bispecific antibody " is herein for referring to the full length antibody generated by following technology: four source hybridomas technology (referring to Milstein et al., Nature, 305 (5934): 537-540 (1983))；Pass through the change of two different monoclonal antibodies Learn conjugation (referring to Staerz et al., Nature, 314 (6012): 628-631 (1985))；Or it is mutated by being introduced in the area Fc Button-enter-hole method or the like is (referring to Holliger et al., Proc.Natl.Acad.Sci.USA, 90 (14): 6444- 6448 (1993)), the method generates a variety of different immunoglobulin substances, and wherein only one is that functional bispecific is anti- Body.Bispecific antibody on one of two combination arm (a pair of of HC/LC) combine a kind of antigen (or epitope), and its second Different antigen (or epitope) is combined on arm (another pair HC/LC).According to this definition, there are two different for bispecific antibody tool Antigen binding arm (at two aspect of specificity and CDR sequence), and be monovalent for its each antigen combined.

" CDR " is herein for referring to " complementary determining region " in antibody variable sequence.In the every of heavy chain and light chain There are three CDR in a variable region.For each variable region, since the N-terminal of heavy chain or light chain, these regions are respectively indicated For " CDR1 ", " CDR2 " and " CDR3 ".Term " CDR group " as used herein refers to that the combination being present in single variable region is anti- One group of former three CDR.Therefore, antigen binding site may include six CDR, and it includes in heavy chain and light chain variable region The CDR group of each.Polypeptide comprising single CDR (such as CDR1, CDR2 or CDR3) is properly termed as " molecular recognition unit " The amino acid residue of verified CDR is formed with the antigen of combination and is contacted extensively the crystallographic analysis of antigen-antibody complex, In widest antigen contact be and heavy chain CDR3.Therefore, molecular recognition unit may be mainly responsible for the spy of antigen binding site It is anisotropic.Generally, CDR residue directly and is most substantially related to influence antigen binding.

The exact boundary of these CDR is subject to difference according to not homologous ray and defines.By Kabat (Kabat et al., Sequences of Proteins of Immunological Interest(National Institutes of Health, Bethesda, Md. (1987) and (1991)) described in system any variable region for being applicable to antibody is not only provided Clear residue numbering system, and provide and define the exact residue boundary of three CDR.These CDR can be described as " Kabat CDR".Chothia and colleague (Chothia and Lesk, J.Mol.Biol., 196:901-917 (1987) and Chothia etc. People, Nature 342:877-883 (1989)) have found that while that there is huge diversity in the level of amino acid sequence, but Certain subdivisions in Kabat CDR use almost the same peptide backbone conformation.These subdivisions be designated as " L1 ", " L2 " and " L3 " or " H1 ", " H2 " and " H3 ", wherein " L " and " H " respectively indicates light chain area and heavy chain region.These regions can be described as " Chothia CDR " has the boundary Chong Die with Kabat CDR.Define other boundaries of the CDR Chong Die with Kabat CDR By Padlan, FASEB J., 9:133-139 (1995) and MacCallum, J.Mol.Biol, 262 (5): 732-745 (1996) it describes.Other CDR boundary definitions may not follow strictly one of system herein, but will be still heavy with Kabat CDR It is folded, but be not significantly affected by the prediction of antigen binding in view of specific residue or residue group or even entire CDR or test and find And can they be shortened or be extended.Method used herein can utilize the CDR defined according to any of these systems, but certain The CDR that a little embodiments are defined using Kabat or Chothia.

" component ", " multiple components " or " at least one component " are often referred to may include for according to side as described herein The examination of method and other method measurements test sample (such as Urine in Patients, whole blood, serum or plasma sample) as known in the art Capture antibody, detectable substance or conjugate, caliberator, control, sensibility group, container, buffer, diluent, salt in agent box, Enzyme, the co-factor of enzyme, detection reagent, pretreating reagent/solution, substrate (for example, as solution), terminate liquid etc..Some components It can in the solution or be lyophilized to be reconstructed and be used in the assay use.

" CT scan " refers to that computerized tomography (CT) is scanned as used herein.CT scan is combined with to be shot from different perspectives A series of radioscopic images, and create using computer disposal the cross sectional image of your internal bone, blood vessel and soft tissue Or slice.X ray CT, positron emission computerized tomography (PET), single photon emission computerized tomography,SPECT can be used in CT scan (SPECT), computer axial direction tomoscan (cat scan) or Computer Aided Tomography.CT scan can be conventional CT and sweep It retouches or spirally/screw type CT scan.In conventional CT scan, slice it is scanned one by one, and in each slice Scanning stops and is moved down into next slice afterwards, for example, being moved down into pelvis at the top of abdomen.Conventional CT scan It is required that patient holds the breath to avoid motion artifacts.Spirally/screw type CT scan is continuous scanning, is clapped in a spiral manner It takes the photograph, and is that wherein scan image is continuous more fast process.

" derivative " of antibody as used herein, which can refer to compared with real or parental antibody, to be had to its amino acid The antibody of one or more modifications of sequence and the domain constructs for showing modification.Derivative still can use day The typical structure configuration of territory found in right antibody, and the amino acid sequence of target (antigen) can be specifically bound.Antibody spreads out The representative instance of biology is the antibody domain or antibody fragment with the antibody of other polypeptides coupling, rearrangement.Derivative can be with Comprising at least one other compound, such as protein domain, the protein domain is connected by covalently or non-covalently key It connects.According to procedures known in the art, connection can be based on genetic fusion.It is present in another in the fusion protein comprising antibody Outer structural domain preferably passes through flexible joint, advantageously peptide linker connection, wherein the peptide linker includes multiple hydrophilic Peptide bonding amino acid, length be enough across other protein domain C-terminal and antibody N-terminal between away from From vice versa.Antibody can be connect with effector molecule, and the effector molecule, which has, is suitable for bioactivity or selective binding example Such as solid support, bioactive substance (such as cell factor or growth hormone), chemical reagent, peptide, protein or drug Conformation.

" being determined by measuring method " determines reference levels or exhausted by any measuring method appropriate for referring to herein To amount.In some embodiments, determine reference levels or absolute magnitude can by with to be administered in the sample from subject The measuring method of measuring method same type realize (such as by immunoassay, the Western Immuno precipitation method, immunoelectrophoresis Method, chemical analysis, SDS-PAGE and western blot analysis or Western Immuno decoration method, electrophoretic analysis, protein Measuring method, competitive binding assay, functional protein measuring method or chromatography or spectroscopic methodology, such as high performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC/MS)).In some embodiments, determine that reference levels or absolute magnitude can pass through It is realized with the measuring method of the measuring method same type to be administered in the sample from subject and under identical determination condition.Such as It is referred to herein go out, present disclose provides (such as the references by comparing different time points of exemplary reference level and absolute magnitude Level calculation).The description provided based on the disclosure is other for those to obtain for other measuring methods modification disclosure herein The measuring method specificity reference levels and absolute magnitude of measuring method are completely in the limit of power of those of ordinary skill in the art. For example, one group of trained sample, including that the sample that obtains from the known human experimenter for suffering from the damage to head is (and more specific Ground suffers from (i) slight TBI from known；Or (ii) moderate to severe TBI human experimenter obtain sample and from it is known not By the damage to head human experimenter obtain sample (more specifically, from it is known not by the mankind of any TBI by The sample that examination person obtains) it can be used for obtaining measuring method specificity reference levels and absolute magnitude.It should be appreciated that " true by measuring method Fixed " and there is the parameter level of cited " sensibility " and/or " specificity " level or absolute magnitude to be used to refer to herein Such reference levels or absolute magnitude, the reference levels or absolute magnitude have been determined when using in the method for the invention The method of cited sensibility and/or specificity is provided.Such as by using multiple and different possibility reference levels and absolutely Sensitivity relevant to reference levels given in the method for the present invention or absolute magnitude is determined to the repetition statistical analysis of the determination data of amount Property and specificity be completely in the limit of power of those of ordinary skill in the art.

" drug abuse " is herein for referring to due to non-medical (effect for such as entertaining and/or changing intelligence) And the one or more additive materials (such as drug) absorbed.Excessively indulge in, use or rely on such drug abuse usually by Referred to as " drug abuse ".The example of drug abuse includes alcohol, barbiturate, benzodiazepine, hemp, can block Cause, psychedelic (such as ketamine, Mosca clever (Pi Yuete (peyote)), PCP, psilocybin, DMT and/or LSD), first quinoline Ketone, opioid drug, amphetamine (including crystal methamphetamine), anabolic steroids, inhalant (contain to have and act on The substance of the volatile materials of the characteristic of spirit, such as nitrite, spray coatings, cleaning solution, marker, glue etc.) and A combination thereof.

" double-specific antibody " is herein for referring to and can tie in each of two combination arm (a pair of of HC/LC) Close the full length antibody (referring to PCT Publication WO 02/02773) of two kinds of not synantigens (or epitope).Therefore, dual specificity combines There are two the same antigen combination arms for having phase homospecificity and identical CDR sequence for albumen tool, and every kind combined for it is anti- It is divalent for original.

" double variable domains " are herein for referring to two or more binding sites on binding protein, the combination Albumen can be (four antigen binding sites) or multivalent binding proteins of (two antigen binding sites) of divalent, tetravalence.DVD It can can be combined in conjunction with a kind of antigen (or a specificity epitope) or polyspecific for monospecific Two or more antigen (i.e. two or more tables of two or more epitopes of same antigen molecule or different target antigens Position) preferably DVD binding protein includes two heavy chain DVD polypeptides and two light chains DVD polypeptide and referred to as " ball is immunized in DVD Albumen " or " DVD-Ig ".Therefore such DVD-Ig binding protein is the tetramer and is similar to IgG molecule, but provide ratio The more antigen binding sites of IgG molecule.Therefore, each half of tetramer DVD-Ig molecule is similar to the half of IgG molecule, It and include heavy chain DVD polypeptide and light chain DVD polypeptide, but a pair of of heavy chain with the offer single antigen binding structural domain of IgG molecule Different with light chain, a pair of of the heavy chain and light chain of DVD-Ig provides two or more antigen binding sites.

The protein-bonded each antigen binding site of DVD-Ig can be originated from donor (" parent ") monoclonal antibody, therefore wrap Heavy-chain variable domains (VH) and light chain containing a total of six CDR for both participating in antigen binding with each antigen binding site can Structure changes domain (VL).Therefore, in conjunction with the DVD-Ig binding protein of two different epitopes, (two of i.e. two different antigen molecules are not With two different epitopes of epitope or same antigen molecule) comprising from the first parental monoclonal antibody antigen binding site and The antigen binding site of second parental monoclonal antibody.

In PCT Publication WO 2007/024715, U.S. Patent number 7,612,181 and Wu et al., Nature Biotech., the description of the design to DVD-Ig binding molecule, expression and characterization is provided in 25:1290-1297 (2007).This The preferred embodiment of class DVD-Ig molecule includes the heavy chain containing structural formula VD1- (X1) n-VD2-C- (X2) n, and wherein VD1 is first Heavy-chain variable domains, VD2 are the second heavy-chain variable domains, and C is heavy chain constant domain, and X1 is that (precondition is connector It is not CH1, and X2 is the area Fc), and n is 0 or 1, but preferably 1；With contain VD1- (X1) n-VD2-C- (X2) n light chain, wherein VD1 is the first light variable domains, and VD2 is the second light variable domains, and C is light chain constant domain, and X1 is that connector is (preceding The condition of mentioning is that it is not CH1, and X2 does not include the area Fc)；And n is 0 or 1, but preferably 1.This DVD-Ig may include two this The heavy chain of sample and two such light chains wherein every chain includes the variable domains being connected in series, and do not have between variable region There is the constant region of intervention, wherein heavy chain and light chain associate to form concatenation function antigen binding site, and a pair of of heavy chain and light Chain can associate with another pair heavy chain and light chain to be formed tool there are four functional antigen binding site tetramer binding protein.In In another example, DVD-Ig molecule may include such heavy chain and light chain, and each heavy chain and light chain include that series connection connects Three variable domains (VD1, VD2, VD3) connect, and there is no the constant region intervened between variable domains, one pair of them weight Chain and light chain can associate to form three antigen binding sites, and one pair of them heavy chain and light chain can with another pair heavy chain and Light chain associate to be formed tool there are six antigen binding site tetramer binding protein.

In a preferred embodiment, DVD-Ig binding protein is not identical only in conjunction with being combined by its parental monoclonal antibody Target molecule, but also the required characteristic with one or more of its one or more parental monoclonal antibody.Preferably, this The other characteristic of kind is the antibody parameter of one or more of parental monoclonal antibody.It can be from its parental monoclonal antibody One or more persons facilitate the protein-bonded antibody parameter of DVD-Ig include but is not limited to antigentic specificity, antigen affinity, effect Power, biological function, epitope identification, protein stability, protein solubility, generation efficiency, immunogenicity, medicine are for power , bioavilability, tissue cross reactivity and ortholog antigen binding.

At least one epitope of DVD-Ig binding protein combination UCH-L1.The protein-bonded non-limiting example packet of DVD-Ig Include the DVD-Ig binding protein of one or more epitopes in conjunction with UCH-L1, epitope and another species (example in conjunction with people UCH-L1 Such as mouse) UCH-L1 epitope DVD-Ig binding protein and combine people UCH-L1 epitope and another target molecule epitope DVD-Ig binding protein.

" dynamic range " refers to the amount of measurement reading with target molecule or analyte in analyzed sample as used herein Proportional band.

It " evaluates " as used herein and " assessment " refers to subject of the assessment with head injury or doubtful head injury To obtain relevant to head injury information, the presence of such as traumatic brain injury is not present and/or degree." evaluation " can be with Level including TBI biomarker (such as UCH-L1) in detection, measurement and/or the quantitative sample from subject.It " comments Valence " can also include that various clinical assessments are carried out to the subject with head injury or doubtful head injury, such as but unlimited In progress GCS assessment, GOSE assessment and/or imaging analysis (such as CT scan or MRI imaging).

" epitope " or " multiple epitopes " or " epitope interested ", which refer to, to be identified on any molecule and it can be combined special The site in the complementary site on specific binding partner.Molecule and specific binding partner are one of specific binding pair Point.For example, epitope can be, in polypeptide, protein, haptens, sugar antigen, (such as, but not limited to glycolipid, glycoprotein or rouge are more Sugar) or polysaccharide.Its specific binding partner can be but not limited to antibody.

" fragment antigen binding fragment " or " Fab segment " refers to the segment of antibody as used herein, simultaneously in conjunction with antigen And a part containing an antigen binding site, complete a light chain and a heavy chain.Fab is by VL, VH, CL and CH1 The monovalent fragment of structural domain composition.Fab by each of heavy chain and light chain a constant domain and a varistructure Domain composition.Variable domains contain paratope (antigen binding site), and it includes one group of complementations of the amino terminal in monomer to determine Determine area.Therefore the epitope on each arm combination antigen of Y.Fab segment can be generated as described in this field, example Such as, using enzyme papain, it can be used for for immunoglobulin monomer being cracked into two Fab segments and Fc segment, or It can be generated by recombination method.

" F (ab') as used herein₂Segment " refers to through the entire IgG antibody of pepsin digestion to remove major part The area Fc, while retaining some hinge areas and the antibody that generates.F(ab')₂Segment has two to link together by disulfide bond The part antigen binding F (ab), therefore be divalent, molecular weight is about 110kDa.Bivalent antibody fragments (F (ab')₂Segment) it is less than It entire IgG molecule and can preferably penetrate into tissue, to promote better antigen recognizing in immunohistochemistry.F (ab')₂The use of segment is also avoided to the Fc receptor or albumin A/G non-specific binding on living cells.F(ab')₂Segment It can be in conjunction with simultaneously Precipitation Antigen.

" frame " (FR) or " Frame sequence " can mean that variable region subtracts the residue sequence of CDR as used herein.Cause Can be determined by not homologous ray (for example, see above) for accurately defining for CDR sequence, thus the meaning of Frame sequence vulnerable to Corresponding different explanations.Six CDR (CDR-L1, CDR-L2 and CDR-L3 of light chain and CDR-H1, CDR-H2 and CDR- of heavy chain H3 the frame on light chain and heavy chain also) is distinguished into four sub-regions (FR1, FR2, FR3 and FR4) on each chain, wherein CDR1 Between FR1 and FR2, CDR2 is between FR2 and FR3, and CDR3 is between FR3 and FR4.Not by specific sub-district In the case where being appointed as FR1, FR2, FR3 or FR4, such as other mentioned framework regions indicate single naturally occurring immune globulin Combination FR in the variable region of white chain.As used herein, FR indicates one in four sub-regions, and FR indicates to constitute frame Two or more in four sub-regions in area.

People's heavy chain and light chain FR sequences are known in the art, and are used as heavy chain and light chain " recipient " frame sequence (or being briefly " recipient " sequence) is arranged to carry out humanizing non-human antibodies by using technology as known in the art.In In one embodiment, people's heavy chain and light chain recipient's sequence are selected from publicly available database such as V-base (hypertext transfer protocol: //vbase.mrc-cpe.cam.ac.uk/) or internationalInformation system (hypertext transfer protocol: // Imgt.cines.fr/texts/IMGTrepertoire/LocusGenes/ the Frame sequence listed in).

" functional antigen binding site " can mean combine on binding protein (such as antibody) as used herein The site of target antigen.The antigen-binding affinity of antigen binding site can be tied not as good as the parent being originated from antigen binding site Hop protein, such as parental antibody are equally strong, but combine antigen ability must be can be used become known for evaluate protein, such as The measurement of any one of antibody and a variety of methods of antigen binding.In addition, multivalent protein herein, such as multivalent antibody The antigen-binding affinity of each antigen binding site is without quantitatively identical.

" Golmud-Lhasa Pipeline " or " GCS " refers to for based on overall sociability or to other as used herein Dependence estimate and 15 subscales of cerebral injury result of classifying.The test is anti-using following values measurement motor reaction, speech Should react with opening eyes: (6- obeys command completely for I, motor reaction；It is positioned when 5- noxious stimulation；4- is retracted from noxious stimulation； 3- exception buckling, i.e. peeling layer state；2- stretching, extension reaction, i.e. decerebrate state；It is reactionless with 1-)；II, verbal response (5- vigilance And it orients；4- speaks confusion, but coherent；3- words is inappropriate and the phrase that is made of words is chaotic；The elusive sound of 2-； There is no sound with 1-；With III, (the spontaneous eye opening of 4- of opening eyes；Eye opening when 3- speaks；It opens eyes when 2- pain；1- does not open eyes).By adding Add the value of I+II+III to determine final scoring.Final scoring can be divided into four possible subsistence levels, lower digital indication More serious damage and worse prognosis: slight (13-15)；(loss of consciousness is more than 30 minutes to moderate disability (9-12)；May or The body or cognitive disorder that may subside: and benefit from rehabilitation)；Severe, which disables (3-8), (goes into a coma: automatism.Do not have The reaction of meaning, without voluntary activity)；It (wake up the period with vegetative state (less than 3)；Awakening, but not with the phase interaction of environment With；There is no orienting response to pain).Moderate traumatic brain injury is defined as leading to the loss of consciousness 20 minutes to 6 hours and Glasgow dusk It is confused the cerebral injury that scale is 9 to 12 points.Patients with severe cerebral injury is defined as that the loss of consciousness is caused to be greater than 6 hours and Glasgow coma amount The cerebral injury that table is 3 to 8.

As used herein, " Glasgow final result scale " refers to the global scale for functional final result, by patient's shape State is assessed as one of following five classifications: death, persistent vegetative state, severe disability, moderate disability restore good.

" Glasgow final result scale of extension " interchangeably used herein or " GOSE " by the way that severe is disabled, moderate The level categories and high-level class that disability and the classification well restored are subdivided into table 1 are classified to provide more detailed eight kinds.

Table 1

Term " humanized antibody " herein for describe comprising from non-human species (such as mouse) heavy chain and gently At least part in chain variable region sequence but wherein VH and/or VL sequence has become more " class people ", i.e., variable with people's system genitale The more like antibody of sequence." humanized antibody " is antibody or its variant, derivative, analog or segment, immunologic specificity Ground combines antigen interested and includes the frame area (FR) of the substantially amino acid sequence with human antibody and substantially with non- The complementary determining region (CDR) of the amino acid sequence of human antibody.As used herein, the term under the background of CDR " substantially " is Refer to amino acid sequence and non-human antibody CDR amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical CDR.Humanized antibody includes substantially all of at least one and usual two varistructures Domain (Fab, Fab', F (ab')₂, FabC, Fv), wherein completely or generally whole CDR regions correspond to non-human immunoglobulin (i.e. Donor antibody) those of CDR region and completely or generally whole framework regions are those of human immunoglobulin(HIg) consensus sequences.In In one embodiment, humanized antibody also includes constant region for immunoglobulin (Fc) (constant region of usual human immunoglobulin(HIg)) At least part.In some embodiments, humanized antibody contains the variable domains of light chain and at least heavy chain.Antibody It also may include the CH1, hinge, the area CH2, CH3 and CH4 of heavy chain.In some embodiments, humanized antibody has contained only source of people Change light chain.In some embodiments, humanized antibody has contained only humanized heavy chain.In certain embodiments, humanization is anti- Body only humanization variable domains containing light chain and/or humanized heavy chain.

Humanized antibody can be selected from any classification of immunoglobulin, including IgM, IgG, IgD, IgA and IgE, and appoint What isotype, including but not limited to IgG1, IgG2, IgG3 and IgG4.Humanized antibody may include from more than one classification Or the sequence of isotype, and it is required to optimize that the specific constant domain of the choice of technology well known in the art can be used Effector function.

The framework region and CDR of humanized antibody are not necessarily to correspond precisely to parental array, such as can be by replacing, being inserted into Or/or lack at least one amino acid residue and to carry out mutagenesis to donor antibody CDR or shared frame the CDR so that at the site Or Framework residues do not correspond to donor antibody or shared frame.However, in a preferred embodiment, such mutation will not be wide General.In general, at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% humanized antibody is residual Base will correspond to those of parent FR and CDR sequence.As used herein, term " shared frame " refers to shared immunoglobulin sequence Framework region in column.As used herein, term " shared immunoglobulin sequences " refers to by associated immunoglobulin sequence family In the sequence that is formed of the most common amino acid (or nucleotide) (see, for example, Winnaker, From Genes to Clones (Verlagsgesellschaft,Weinheim,1987)).Therefore, " shared immunoglobulin sequences " may include " shared frame Frame area " and/or " shared CDR ".In immunoglobulin class, each position in consensus sequence in family by most often appearing in The amino acid of that position occupies.It may include any one in consensus sequence if two amino acid continually occur on an equal basis.

As this paper under the background of two or more polypeptides or polynucleotide sequence used in it is " identical " or " same Property " it can refer to sequence identical residue with prescribed percentage on specified region.The percentage can be calculated by following: It most preferably compares two sequences, compare two sequences, the position for determining residue identical in the two sequences in specified region Quantity to generate the quantity of matching position, the total quantity with the quantity of matching position divided by the position in specified region, and Result is generated to the percentage of sequence identity multiplied by 100.Two sequences have different length or compare generate one or In the case that multiple staggered ends and the specified region compared only include single sequence, the residue of single sequence is included in calculating Denominator rather than in molecule.

As " damage to head " interchangeably used herein or " head injury " refer to scalp, skull or brain Any wound.Such damage may include the only slight shock on skull or may be serious cerebral injury.Such damage packet Include the primary injury of brain and/or the secondary lesion of brain.Primary brain occurs during initial infringement, and Caused by the displacement of the physical structure of brain.More specifically, primary brain is occurred during traumatic event to essence The physical injury of (tissue, blood vessel) leads to shearing and compressing to surrouding brain tissue.Secondary brain injury occurs to damage in primary After wound, and it may relate to a series of cell processes.More specifically, secondary brain injury refers to after primary brain The interior variation developed (from a few hours to a couple of days) for a period of time.It includes facilitating in the brain for further destroying brain tissue carefully The entire cascade of born of the same parents, chemistry, tissue or Vascular change.

Closed or open (penetrability) can be to the damage on head.Closed head injury refers to wherein Skull is not knocked the wound of the scalp of object penetration, skull or brain.Open head injury refers to that wherein skull is hit Hit the scalp of object penetration, the wound of skull or brain.The injury on head, which can be, is shaken, by the body of people by leading to closed Or external mechanical or other power of open head trauma are (for example, the traffic accident when such as automobile, aircraft, train； Fiercely attack on head such as with baseball bat or from firearms) generate passivity impact, cerebrovascular accident (such as apoplexy), once or it is more It is secondary fall (for example, as movement or other activities), explosion or shock wave (being referred to as " blast injury ") and by other types of Caused by blunt forces wound.It optionally, may be by taking in and/or being exposed to chemical substance, toxin or chemicals to the damage on head The combination of matter and toxin causes.Such chemical substance and/or the example of toxin include fire, mould, asbestos, insecticide and insecticidal Agent, organic solvent, paint, glue, gas (such as carbon monoxide, hydrogen sulfide and cyanide), organic metal (such as methyl mercury, Lead tetraethide and organotin) and/or one or more drug abuse.It optionally, may be due to subject to the damage on head Suffer from autoimmune disease, metabolic disorder, brain tumor, one or more viruses, meningitis, hydrocephalus, anoxic or its any group Caused by conjunction.In some cases, can not determine whether that any such event or damage occurred.For example, patient or subject May there is no medical history, subject possibly can not speak, and subject may realize that the event etc. that they are exposed to.Such situation It is described herein as subject " injury that head may be suffered from ".In the certain embodiments of this paper, closed head Damage does not include and particularly excludes cerebrovascular accident, such as apoplexy.

As used herein " isolated polynucleotides " can mean polynucleotides (such as genome, cDNA or synthesis come The polynucleotides in source or combinations thereof), according to its source, the polynucleotides are not seen in nature with " isolated polynucleotide " All or part of association of locating polynucleotides；It is operably coupled to its unconnected polynucleotides in nature； Or it is present in nature not as a part of larger sequence.

It " marks " as used herein and " detectable label " refers to and be attached to antibody or analyte, so that antibody and analysis The detectable part of reaction between object, and the antibody or analyte so marked is referred to as " detectably marking ".Mark Note can produce the signal that can be detected by vision or instrument means.Various labels include the substance for generating signal, are such as sent out Color group, fluorescent chemicals, chemiluminescence compound, radioactive compound etc..The representative example of label includes generating the portion of light Point, such as acridine compounds, and generate the part of fluorescence, such as fluorescein.This document describes other labels.In this respect, Described part itself can be undetectable, but can become detectable when reacting with another part.Term is " detectably Label " use be intended to cover this label.

Any suitable detectable label as known in the art can be used.For example, detectable label can be radioactivity Mark (such as 3H, 14C, 32P, 33P, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho and 153Sm), enzyme mark Remember (horseradish peroxidase, alkaline peroxidase, G 6 PD etc.), chemiluminescent labeling (such as acridine Ester, thioesters or sulfonamide；Luminol, different luminol, phenanthridines ester etc.), fluorescent marker (such as fluorescein (such as 5- fluorescein, 6- Fluoresceincarboxylic acid, 3'6- Fluoresceincarboxylic acid, 5 (6)-Fluoresceincarboxylic acid, 6- chlordene-fluorescein, 6- tetrachlorofluorescein, different sulphur cyanogen Sour fluorescein etc.)), rhodamine, phycobniliprotein, R- rhodophyll, the quantum dot cadmium selenide of capping (such as zinc sulphide), thermometric label Or Immuno polymerase chain reaction label.The introduction of label, label program and label detection sees Polak and Van Noorden, Introduction to Immunocytochemistry, second edition, Springer Verlag, N.Y. (1997)；With Haugland, Handbook of Fluorescent Probes and Research Chemicals (1996) (its be by Combination handbook and catalogue published by Molecular Probes, Inc., Eugene, Oregon) in.Fluorescent marker can be used for FPIA In (see, for example, U.S. Patent number 5,593,896,5,573,904,5,496,925,5,359,093 and 5,352,803, lead to It crosses reference to be integrally incorporated herein).Acridine compounds can be used as in homogeneous chemiluminescent measuring method detectable label (see, for example, Adamczyk et al., Bioorg.Med Ghem.Lett.16:1324-1328 (2006)；Adamczyk et al., Bioorg.Med Chem.Lett.4:2313-2317(2004)；Adamczyk et al., Biorg.Med Chem.Lett.14:3917-3921 (2004)；With Adamczyk et al., Org.Lett.5:3779-3782 (2003)).

In one aspect, acridine compounds are acridine -9- formamides.The method for being used to prepare acridine 9- formamide is described in Mattingly,J.Biolumin.Chemilumin.6:107-114(1991)；Adamczyk et al., J.Org.Chem.63: 5636-5639(1998)；Adamczyk et al., Tetrahedron 55:10899-10914 (1999)；Adamczyk et al., Org.Lett.1:779-781(1999)；Adamczyk et al., Bioconjugate Chem.11:714-724 (2000)； Mattingly et al., In Luminescence Biotechnology:Instruments and Applications； Dyke, K.V. are edited；CRC Press:Boca Raton, the 77-105 pages (2002)；Adamczyk et al., Org.Lett.5: 3779-3782(2003)；(it is whole each by reference with U.S. Patent number 5,468,646,5,543,524 and 5,783,699 It is incorporated herein for it in the introduction about this aspect).

Another example of acridine compounds is acridine -9- aryl ester carboxylic acid.Acridine -9- aryl ester carboxylic acid with Formula II An example be 10- methyl -9- (phenyloxycarbonyl) acridine fluosulfonic acid ester (be purchased from Cayman Chemical, Ann Arbor, MI).The method for being used to prepare acridine -9- aryl ester carboxylic acid is described in McCapra et al., Photochem.Photobiol.4: 1111-21(1965)；Razavi et al., Luminescence 15:245-249 (2000)；Razavi et al., Luminescence 15:239-244(2000)；(it is integrally incorporated this each by reference with U.S. Patent number 5,241,070 Text is for it in the introduction about this aspect) in.Such acridine -9- aryl ester carboxylic acid is for by least one oxidizing ferment oxygen Change in analyte the hydrogen peroxide that the generates efficient chemiluminescent indicator in terms of signal strength and/or signal rapidly degree. The chemiluminescence emission process of acridine -9- aryl ester carboxylic acid quickly completes, i.e., completes in 1 second, and acridine -9- formamide chemistry Luminescence emissions are extended to 2 seconds.However, acridine -9- aryl ester carboxylic acid loses its chemiluminescent properties in the presence of protein.Cause This, using needing during signal generates and detects, there is no protein.For separating or removing the side of protein in sample Method is well known to those skilled in the art, and including but not limited to ultrafiltration, extraction, precipitating, dialysis, chromatography and/or digestion (see, for example, Wells, High Throughput Bioanalytical Sample Preparation.Methods and Automation Strategies,Elsevier(2003)).The amount of removal or isolated protein can be with from test sample About 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90% or about 95%.More details about acridine -9- aryl ester carboxylic acid and application thereof are set forth in what on April 9th, 2007 submitted In U.S. Patent number 11/697,835.Acridine -9- aryl ester carboxylic acid is soluble in any suitable solvent, is such as deaerated Anhydrous N,N-dimethylformamide (DMF) or aqueous sodium taurocholate.

" catenation sequence " or " connection peptide sequence " refers to and one or more polypeptide of interest sequences (such as overall length, segment Deng) connection natural or artificial polypeptide sequence.Term " connection " refers to the engagement of catenation sequence Yu polypeptide of interest sequence.This Class polypeptide sequence preferably passes through one or more peptide keyed engagements.Catenation sequence can have the length of about 4 to about 50 amino acid Degree.Preferably, the length of catenation sequence is about 6 to about 30 amino acid.It can be repaired by amino acid replacement, adding or deletion Natural catenation sequence is adornd to generate artificial connection sequence.Catenation sequence can be used for many purposes, including in recombinant Fab.Show Example property catenation sequence includes but is not limited to: (i) histidine (His) label, such as 6X His label, with HHHHHH (SEQ ID NO:2) amino acid sequence, can be used as catenation sequence to be conducive to separate and purify polypeptide of interest and antibody；(ii) intestines Kinase lysis site, such as His label, for separating and purifying interested protein and antibody.Usually, by Enterokinase cleavage position Point is used to separate and purify interested protein and antibody together with His label.Various enterokinase cleavage sites are in this field In be known.The example of enterokinase cleavage site include but is not limited to DDDDK (SEQ ID NO:3) amino acid sequence and its Derivative (such as ADDDDK (SEQ ID NO:4) etc.)；(iii) miscellaneous sequence can be used for linking or connecting single chain variable fragment Light chain and/or heavy chain variable region.The example of other catenation sequences is found in Bird et al., Science 242:423-426 (1988)；Huston et al., PNAS USA 85:5879-5883 (1988)；With McCafferty et al., Nature 348: In 552-554 (1990).Catenation sequence can also be modified with for other function, the attachment of such as drug or attach to solid Support.In the context of the disclosure, monoclonal antibody can for example contain catenation sequence, and such as His label, enterokinase are split Solve site or both.

" monoclonal antibody " refers to the antibody that the group of basically homogeneous antibody obtains as used herein, i.e., except can be with Except a small amount of existing possible naturally occurring mutation, it is identical for constituting the individual antibody of the group.Monoclonal antibody needle There is high degree of specificity to single antigen.In addition, more grams from the different antibodies for generally including to be directed to different determinants (epitope) Grand antibody preparation is different, and every kind of monoclonal antibody is for the single determinant on antigen.The monoclonal antibody of this paper is specifically wrapped Include " chimeric " antibody, wherein a part of heavy chain and/or light chain be originated from particular species or belong to specific antibodies classification or subclass Antibody in corresponding sequence it is identical or homologous, and the remainder of the chain be originated from another species or belong to another antibody class Corresponding sequence in the segment of other or subclass antibody and this kind of antibody is identical or homologous, as long as they show required life Object activity.

" MRI " used herein refers to magnetic resonance imaging, is the body being used to form in health and disease in radiology The medical imaging technology of the picture of the anatomical structure and physiology course of body.MRI scanner based on nuclear magnetic resonance (NMR) science makes The image of body interior is generated with high-intensity magnetic field, radio wave and field gradient.

Term " multivalent binding proteins " is herein for referring to comprising two or more antigen binding sites (herein Also referred to as " antigen-binding domains ") binding protein.Multivalent binding proteins are preferably engineered to have three or more A antigen binding site, and not usually naturally occurring antibody.Term " multi-specific binding protein " refers to and can combine The binding protein of two or more related or uncorrelated targets, including can combine identical target molecule two or more not With the binding protein of epitope.

As when " negative predictive value " or " NPV " interchangeably used herein, which refers to, assumes that they have negative test result Subject has the probability of negative final result.

" reference levels " refer to for assessing diagnosis, prognosis or the measurement critical value of therapeutic efficiency, In as used herein Herein by it with various clinical parameters (such as the presence of disease, disease stage, disease severity, the progress of disease, not into Exhibition or improvement etc.) it connects or is associated.As used herein " absolute magnitude " refer in different time points obtain or sample The absolute value of variation or difference between at least two measurement results, and it is similar with reference levels, by itself and various clinics Parameter (such as disease presence, disease stage, disease severity, the progress of disease, be not in progress or improve) connect or It is associated." absolute value " refers to that (such as two comparison levels (such as are obtained in first time point real number as used herein It is horizontal in the level that the second time point obtained)) between difference size, without considering its symbol, i.e., no matter it is just Or it is negative.

Present disclose provides exemplary reference level and absolute magnitude (such as the reference levels meters by comparing different time points It calculates).It is well known, however, that reference levels and absolute magnitude can according to the property of immunoassay (such as using antibody, anti- Answer condition, sample purity etc.) and change, and can compare and standardized assay.The description provided based on the disclosure is directed to Other immunoassay modification disclosure hereins are to obtain the immunoassay specificity ginseng for those other immunoassays Examining horizontal and absolute magnitude is further completely in the limit of power of those of ordinary skill in the art.Although reference levels and The exact value of absolute magnitude can change between measuring method, but discovery as described herein should be blanket and can be outer It is pushed into other measuring methods.For example, the bigger time resolution of the UCH-L1 level under the background of data offer TBI provided herein Rate, and it is not limited to single time point assessment.Data from the disclosure prove UCH-L1 level dynamic change after injury, this A variety of different measurements can be used to evaluate.

" point-of-care device " refer at or near point-of-care (i.e. outside laboratory), patient care when Between and place (such as in hospital, doctor's office, urgent or other medical care institutions, Huan Zhejia, home for destitute and/or long-term In nursing and/or deathbed care facility) device of medical diagnosis test is provided.The example of point-of-care device includes by Abbott Device (such as i-STAT and i-STAT Alinity, the pervasive biology of Laboratories (Abbott Park, IL) production Sensor (Rowville, Australia) (referring to US 2006/0134713), Axis-Shield PoC AS (Oslo, ) and clinical labororatory's product (Los Angeles, USA) Norway.

As when " positive predictive value " or " PPV " interchangeably used herein, which refers to, assumes that they have positive test result Subject has the probability of positive final result.

" quality control reagents " under immunoassay as described herein and the background of kit include but is not limited to school Quasi- object, reference material and sensibility group.Come usually using " caliberator " or " reference substance " (such as one or more, such as plural number kind) Correction (standard) curve is established with the concentration of interpolation analyte (such as antibody or analyte).It is alternatively possible to use close ginseng Examine the single caliberator of horizontal or control level (such as " low ", " medium " or "high" are horizontal).A variety of caliberators can be used in combination (i.e. more than one caliberator or different amounts of caliberator) is with composition " sensibility group ".

" recipient's operating characteristics " curve or " ROC " curve refer to illustrate binary classification system its discrimination threshold variation when The figure of performance is drawn.For example, ROC curve is the true positive rate of cut off possible for the difference of diagnostic test compared to false sun The drawing of property rate.It is by various threshold values setting under by the true-positive fraction (TPR=true positive rate) in the positive relative to False-positive fraction (FPR=false positive rate) in feminine gender carries out drawing generation.TPR is also referred to as sensibility, and FPR is one to subtract Go specificity or true negative rate.ROC curve illustrates tradeoff (any increase all companions of sensibility between sensitivity and specificity With the reduction of specificity)；Curve gets over the left margin closely along the space ROC, followed by top boundary, and it is more accurate to test； Curve is tested more inaccurate closer to 45 degree of diagonal lines in the space ROC；Tangent slope at cut off provides the test value seemingly Right rate (LR)；And area under the curve is the measurement of text accuracy.

" recombinant antibodies " and " a variety of recombinant antibodies " refer to the antibody prepared by one or more steps, including pass through weight The all or part of nucleic acid sequence for encoding one or more monoclonal antibodies is cloned into expression vector appropriate by group technology, And antibody then is expressed in host cell appropriate.The term includes but is not limited to recombinate the monoclonal antibody, embedding generated It closes antibody, humanized antibody (all or part of humanization), the more characteristics formed by antibody fragment or multivalent structure, difunctional resist Body, special-shaped conjugation Ab, DVD-With other antibody described in (i) herein.(double variable domains immunoglobulins and its system Preparation Method is described in Wu, C et al., in Nature Biotechnology, 25:1290-1297 (2007)).It is used herein Term " bifunctional antibody " refer to including with the specificity for antigen site the first arm with have for Bu Tong anti- The antibody of second arm of the specificity put in situ, i.e. bifunctional antibody have dual specificity.

As used herein to " risk assessment " of subject (such as patient), " classification of risks ", " risk identification " or " wind Danger layering " refers to that evaluation includes the factor of biomarker, includes that seizure of disease or the future event of progression of disease are sent out with prediction Raw risk, so as to make the Treatment decsion about subject on the basis of more knowing.

" sample ", " test sample ", " sample ", " sample from subject ", " Patient Sample A " and " biological sample " can To be used interchangeably and can be blood sample (such as whole blood), tissue, urine, serum, blood plasma, amniotic fluid, myelencephalon herein Liquid, placenta cells or tissue, endothelial cell, leucocyte or monocyte.Sample such as can be used directly from patient with obtaining, or Person can pre-process, and by filtering, dilution, extraction, concentration, centrifugation, inactivated to interfering component, add reagent etc., from And by it is described herein or it is other it is more known in the art in a manner of modify the feature of sample.In one embodiment, Sample is whole blood sample, serum or plasma sample.In another embodiment, sample is whole blood.In another embodiment, sample Product are serum.In another embodiment again, sample is blood plasma.In other embodiment again, sample is obtained from people Whole blood sample.In still another embodiment, sample is the plasma sample obtained from people.Going back another embodiment again In, sample is the blood serum sample obtained from people.

Various kinds of cell type, tissue or body fluid be can use to obtain sample.Such cell type, tissue and fluid can be with Including histotomy, such as biopsy and autopsy samples, obtain the frozen section for histology purpose, blood (such as whole blood), Blood plasma, serum, red blood cell, blood platelet, interstitial fluid, celiolymph etc..Cell type and tissue can also include lymph, brain ridge Marrow liquid, the fluid collected by tissue or cell type can be provided by removing cell sample from the mankind and non-human animal, But can also by using be previously separated cell (such as separated by other people, at other times, and/or for other Purpose) it completes.Also filing tissue can be used, such as with those for the treatment of or final result history.May not be needed protein or Nucleotide separation and/or purifying.

" sensibility " refers to that final result is correctly to be accredited as positive (such as correctly to identify that those are suffered from positive subject The subject of their just tested diseases or medical condition) ratio.For example, this may include subject from slight Moderate is correctly accredited as in those of TBI to severe TBI, by subject from correct in those of moderate to severe TBI Ground is accredited as TBI is never suffered from slight TBI, by subject in those of be accredited as with moderate to severe TBI or will It is correctly accredited as in those of subject never TBI with slight TBI).

" specificity " of measuring method as used herein refers to that final result is correctly to be accredited as feminine gender in negative subject The ratio of (such as correctly identifying that those do not suffer from the subject of their just tested diseases or medical condition).For example, this can Can include by subject from be correctly accredited as in those of slight TBI do not suffer from moderate to severe TBI, by subject from It is not suffering from being correctly accredited as not suffering from slight TBI or be correctly accredited as subject in those of moderate to severe TBI There is any TBI).

" calibration composition series " refers to the numerous compositions of the UCH-L1 comprising known concentration, wherein every kind of composition with It is described series in other compositions the difference is that UCH-L1 concentration.

" Single Molecule Detection " refers to low-down concentration level as used herein (such as pg/mL or fg/mL are horizontal) The individual molecule of lower detection and/or the analyte in measurement test sample.Many different unimolecular analyzers or device are at this It is known in field and including nano-pore and nanometer well device.The example of nanopore device is described in International Patent Publication No. In WO 2016/161402, it is incorporated hereby accordingly.The example of nanometer well device is described in international monopoly public affairs In the number of opening WO 2016/161400, it is incorporated hereby accordingly.

It can be used for being attached and/or attracting and fix as " solid phase " or " solid support " interchangeably used herein refers to Change (1) one or more capturing agents or capture specific binding partner, or (2) one or more detection agents or detection specificity Any material of binding partners.Solid phase can attract with regard to it and the capability of immobilization capturing agent is selected.Optionally, Solid phase can have the bridging agent adhered on it, and the bridging agent, which has, to be attracted with immobilization (1) capturing agent or capture specifically Property binding partners, (2) detection agent or detection specific binding partner.For example, bridging agent may include charge species, phase For capturing agent (such as capture specific binding partner) or detection agent (such as detection specific binding partner) itself or It is conjugated relative to (1) capturing agent or capture specific binding partner, or (2) detection agent or detection specific binding partner Charge species be oppositely charged.In general, bridging agent can be any binding partners (preferably anisotropic), It is immobilized in and (is attached to) in solid phase and have and matched by association reaction come immobilization (1) capturing agent or capture specific binding Even body, or (2) detection agent or the ability for detecting specific binding partner.Bridging agent make capturing agent before performance measurement or During performance measurement indirectly in conjunction with solid phase material.For example, solid phase can be plastics, derivative plastics, magnetism or non magnetic gold Category, glass or silicon, including such as test tube, microtiter well, thin slice, bead, particle, chip and those of ordinary skill in the art are The other constructions known.

It " specifically binds " as used herein or " specifically combining " can refer to antibody, protein or peptide and The interaction of two chemical substances, wherein interaction is dependent on specific structure in chemical substance (such as antigenic determinant or table Position) presence；For example, antibody identifies and combines specific protein structure, rather than broad incorporation protein.If antibody pair Epitope " A " be it is specific, then in containing labeled " A " and the reaction of antibody, the molecule of the A containing epitope (or it is free not Label A) presence will reduce the amount of the labeled A in conjunction with antibody.

" specific binding partner " is the member of specific binding pair.Specific binding to comprising two different moleculars, It by chemically or physically specifically binding each other.Therefore, except the antigen of common immunoassay and antibody specificity knot It closes to except, other specific bindings are to may include biotin and avidin (or streptavidin)；Carbon water Compound and agglutinin；Complementary nucleotide sequence；Effector molecule and acceptor molecule；Co-factor and enzyme；Enzyme and enzyme inhibitor etc..This Outside, specifically bind to may include be original specific binding members analog member, such as analyte-analog.Exempt from Epidemic disease reactivity specific binding members include separation or recombination generate antigen, antigen fragment and antibody, including monoclonal and Polyclonal antibody and its compound and segment.

" statistically significant " refers to the relationship between two or more variables by except random machine as used herein A possibility that other factors except meeting cause.Assumed statistical inspection is used to determine whether the result of data set to have statistics On conspicuousness.In assumed statistical inspection, as long as the p value for the test statistics observed is less than the conspicuousness water of research definition It is flat, just obtain significance,statistical result.P value, which assumes that, obtains the pole at least as the result observed when null hypothesis is true The probability of the result at end.The example of statistical hypothesis analysis includes the inspection of Wilcoxon symbol rank, t inspection, card side or Fei Sheer It is accurate to examine." conspicuousness ", which refers to, as used herein is not yet determined as having the variation of significance,statistical (such as it can Assumed statistical inspection can not be undergone).

" subject " and " patient " is interchangeably used for referring to any vertebrate as used herein, including but not limited to feeds Newborn animal (such as it is ox, pig, camel, yamma, horse, goat, rabbit, sheep, hamster, cavy, cat, dog, rat and mouse, inhuman Primate (such as monkey, machin or rhesus macaque, chimpanzee etc.) and the mankind).In some embodiments, tested Person can be the mankind or non-human.In one embodiment, class that subject is a human.Subject or patient can undergo other shapes The treatment of formula.

" treatment (Treat/treating/treatment) " respectively herein it is interchangeable for describe to reverse, mitigate or The progress of the disease and/or damage that inhibit this term to be applicable in or one or more symptoms of this disease.According to subject Symptom, the term also refers to prevention disease, and the breaking-out including preventing disease or prevention symptom relevant to disease.It controls Treatment can be carried out in a manner of acute or chronic.The term also refers to that reduction is relevant to this disease before being tormented by disease The seriousness of disease or symptom.This prevention disease or reduction disease severity before torment refer to not to torment by disease Administration time pharmaceutical composition is applied to subject." prevention " also refers to prevention disease or one kind relevant to this disease Or the recurrence of a variety of symptoms." treatment " and the behavior for " therapeutically " referring to treatment, as " treatment " is as defined above.

Refer to the complicated damage with wide spectrum symptom and disability such as " traumatic brain injury " or " TBI " used interchangeably herein Wound.TBI is many times analogous to the acute events of other damages.TBI can be divided into " slight ", " moderate " or " severe ".TBI's The reason is that diversified, and including the shake of the body of such as people, traffic accident, firearms damage, cerebrovascular accident (such as apoplexy), Tumble, explosion or shock wave and other types of blunt forces wound.Other reasons of TBI include taking in and/or being exposed to one kind Or multi-chemical or toxin (such as fire, mould, asbestos, insecticide and insecticide, organic solvent, paint, glue, gas (such as carbon monoxide, hydrogen sulfide and cyanide), organic metal (such as methyl mercury, lead tetraethide and organotin), one kind or more Kind drug abuse or combinations thereof).Optionally, TBI may suffer from autoimmune disease, metabolic disorder, brain tumor, one kind or more Occur in the subject of kind virus, meningitis, hydrocephalus, anoxic or any combination thereof.Young people and the elderly be TBI risk most High age group.In the certain embodiments of this paper, traumatic brain injury or TBI do not include and particularly exclude the cerebrovascular It is unexpected, such as apoplexy.

" slight TBI " refers to cerebral injury as used herein, wherein the loss of consciousness be of short duration and usually several seconds or A few minutes and/or chaotic and disorientation are shorter than 1 hour.Slight TBI is also referred to as cerebral concussion, slight head trauma, light Micro- TBI, minimal brain damage and slight head injury.Although MRI and CT scan are often normal, with slight TBI Body may have a cognitive question, such as headache, thinking difficulty, memory problems, attention deficit, mood swing and dejected.

Slight TBI is most common TBI, and is usually missed in initial damage.In general, subject has in 13- The Golmud-Lhasa Pipeline number of (such as 13-15 or 14-15) between 15.1 15 (15%) slight TBI patient's Symptoms last 3 months or longer time.Slight TBI is defined as the forceful motion on head or leads to the state of mind less than 30 minutes The result of the impact force of of short duration variation (chaotic, disorientation or the loss of memory) or the loss of consciousness.The common sympton packet of slight TBI Include that fatigue, headache, vision disorder, memory loss, attention/concentrated force be poor, sleep disturbance, dizziness/disequilibrium, irritability Emotional handicap, depressed mood and epilepsy.Other symptoms relevant to slight TBI include nausea, anosmia, to light and sound Sensibility, emotional change, vast and hazy or chaotic, and/or thought slowness.

As used herein, " moderate TBI " refers to cerebral injury, and wherein the loss of consciousness and/or chaotic and disorientation are 1 to 24 Between hour and subject has the Golmud-Lhasa Pipeline number between 9-13 (such as 9-12 or 9-13).With moderate The individual of TBI has abnormal Brian Imaging result." severe TBI " refers to cerebral injury as used herein, and wherein the loss of consciousness is super Spend 24 hours and the loss of memory after damage or penetrability head injury grew 24 hours and subject have 3-8 between Golmud-Lhasa Pipeline number.The range of defect is from the Cognitive function damage of higher level to comatose state.Survivor may Abnormal, phrenoplegia or emotional problem with limited arm or leg function, speech or language.With severe injury Individual may be chronically at state of anergy.For many people with severe TBI, it usually needs long-term rehabilitation is with most It functions to limits and independence.

The common sympton of moderate to severe TBI include cognitive defect, including attention, concentrated force, distractibility, note Recall the difficulty in terms of power, arithmetic speed, confusion, perseveration of speech, impulsion, Language Processing and/or " executing function ", understand spoken language Word (sensory aphasia), speak and be understood difficult (logaphasia), glossolalia, speech rate quickly or it is very slow, Reading problem, writing, explain touchs, temperature, movement, position and it is fine distinguish difficult, sense impression is integrated or Medelling in pairs significant data, part or all of visual loss, eye muscle inability and the double vision (diplopia) of psychology, eye-blurred, Judge apart from the problem of, involuntary eye movement (nystagmus), intolerant to light (photophobia), hearing (such as hearing weaken or Lose, there is song (tinnitus) in ear, the susceptibility of sound increased), anosmia or weaken (anosmia), ageusia Or weaken, convulsions relevant to epilepsy, the convulsions may be several types and may relate to consciousness, sensory perception or movement Flesh is mobile, interrupts to the control of intestines and bladder, insomnia, endurance forfeiture, appetite change, body heat regulation, difficult menstruation, dependency line For, changeable in mood ability, power shortage, irritability, aggressiveness, depression ,/shortage consciousness of disinthibiting or refusal.

As " ubiquitin carboxy terminal hydrolase-l 1 " or " UCH-L1 " used interchangeably herein refers in the mankind by UCH-L1 The deubiquitinating enzymes of gene coding.UCH-L1 is also referred to as ubiquitin carboxy terminal esterase L1 and ubiquitin thioesterase, is the hydrolysis of its product The small C-terminal adduct of ubiquitin is to generate the member of the gene family of ubiquitin monomer.

The level or amount that " UCH-L1 state " can mean UCH-L1 at some time point are (such as using the list of UCH-L1 A measured value), the level of UCH-L1 relevant to monitoring or amount (retest such as is carried out to identify UCH-L1 amount to subject Increase or decrease), it is treatment-related with traumatic brain injury (either primary brain and/or secondary brain injury) Level or amount of UCH-L1 or combinations thereof.

" variant " is herein for describing due to the insertion of amino acid, missing or conservative replaces in amino acid sequence side Face is different, but retains the peptide or polypeptide of at least one bioactivity.The representative example of " bioactivity " includes anti-by specificity Body combines or promotes the ability of immune response.Variant is also substantially the same with reference protein for describing to have herein The protein of amino acid sequence, the reference protein have the amino acid sequence for retaining at least one bioactivity.Amino acid Conservative replaces, i.e., replaced with the different aminoacids of similar characteristic (such as hydrophily, the degree of charging zone and distribution) Amino acid is acknowledged as being usually directed to minor change in the art.As understood in the art, these minor changes can be with It is identified by considering the hydrophilic and hydrophobic index of amino acid part.Kyte et al., J.Mol.Biol.157:105-132 (1982). The hydrophilic and hydrophobic index of amino acid is based on the considerations of its hydrophobicity and charge.It is known in the art that similar hydrophobicity refers to Several amino acid can be substituted and still retaining protein function.In one aspect, the amino acid that hydrophobicity index is ± 2 is taken Generation.The hydrophily of amino acid, which can also be used to disclose, will generate the substitution for the protein for retaining biological function.It is examined under the background of peptide The hydrophily for considering amino acid allows to calculate the maximum local average hydrophilicity of the peptide, be it is a kind of be reported with antigenicity and The good associated useful measurement of immunogenicity.U.S. Patent number 4,554,101 is hereby incorporated by reference in their entirety.Such as ability Understood in domain, the substitution of the amino acid with similar hydrophilicity score, which can produce, retains bioactivity (such as immunogenicity) Peptide.It can be replaced with the amino acid with the hydrophilicity value in ± 2 each other.The hydrophobicity index and hydrophily of amino acid Both value is influenced by the specific side chain of the amino acid.It is the amino acid compatible with biological function that observation result is consistent with this Replace be understood to depend on amino acid and especially those amino acid side chain relative similarities, such as by hydrophobicity, Hydrophily, charge, size and other characteristics are revealed." variant " can also be used for referring to the antigen reactivity of anti-UCH-L1 antibody Segment, it is different from the respective segments of anti-UCH-L1 antibody in terms of amino acid sequence, but still there is antigen reactivity and can To be competed with the respective segments for the anti-UCH-L1 antibody in conjunction with UCH-L1." variant " can also be used for description differentially Process (such as passing through proteolysis, phosphorylation or other posttranslational modifications), but still retain its antigen reactivity polypeptide or Its segment.

" carrier " is used to describe to transport the nucleic acid molecules for another nucleic acid that it has connected herein.It is a type of Carrier is " plasmid ", and the plasmid, which refers to, can be connected to additional DNA section circular double stranded DNA ring therein.It is another The carrier of type is viral vectors, wherein additional DNA section may be connected in viral genome.Certain carriers can be Independently replicated in the host cell that they are introduced into (such as bacteria carrier and episomal mammalian comprising bacterial origin of replication Carrier).It is thin that other carriers (such as non-add type mammalian vector) can be integrated into host later in being introduced into host cell In the genome of born of the same parents, and thus replicated together with host genome.In addition, certain carriers can instruct their institutes operationally The expression of the gene of connection.Examples of such carriers is referred to herein as " recombinant expression carrier " (or being referred to as " expression vector ").Generally For, the expression vector suitable for recombinant DNA technology is usually in plasmid form." plasmid " and " carrier " is used interchangeably, because It is most common carrier format for plasmid.However, it is possible to use the expression vector of the other forms of identical functions is played, it is such as viral Carrier (such as replication defect type retrovirus, adenovirus and adeno-associated virus).In this regard, the RNA pattern of carrier (including rna virus vector) can be used in the context of the disclosure.

Unless being defined otherwise herein, the scientific and technical terms for otherwise disclosure being combined to use will have common by this field The meaning that technical staff is generally understood.For example, in conjunction with cell and tissue culture described herein, molecular biology, being immunized Any nomenclature and its technology that, microbiology, science of heredity and protein and nucleic acid chemistry and hybridization use are abilities It is known and those of common in domain.The meaning and scope of term should be clearly；However, if there is any implicit ambiguity, Then definition presented herein is prior to any dictionary or external definition.In addition, unless additionally by context demands, otherwise The term of odd number should include plural number and plural term should include odd number.

2, the method whether assisted diagnosis and evaluation human experimenter suffer from the damage to head

In addition to other methods, this disclosure relates to which whether a kind of assisted diagnosis and evaluation human experimenter suffer from or may be By the method for the damage to head.The method can have the tested of the doubtful damage to head with assisted diagnosis and evaluation Person traumatic brain injury degree (such as determine subject whether suffer from slight TBI or moderate to severe TBI).Such as this paper institute With " determining whether subject suffers from mild trauma cerebral injury or moderate to severe trauma cerebral injury ", which refers to, uses the side Method (such as using the other information of such as clinical assessment data) determines that subject is more likely to slight TBI or moderate to weight Spend TBI.The method may include sample is obtained in 24 hours to subject's suspicious lesion, make sample with for traumatic (the early stage biomarker of the traumatic brain injury is by ubiquitin carboxy terminal hydrolase-l 1 for the early stage biomarker of cerebral injury (UCH-L1) form) antibody contact, to allow to be formed the compound of antibody and UCH-L1.In alternative, the side Method may include obtain the first sample obtained from human experimenter in first time point and the second time point obtain second Sample, the first sample is derived from human experimenter in 24 hours of damage, and the second sample is after first time point Second time point was derived from human experimenter in such as about 3 hours to about 6 hours after obtaining the first sample；By the first sample and Two samples respectively be directed to traumatic brain injury early stage biomarker (the early stage biomarker of the traumatic brain injury Be made of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1)) antibody contact, to allow to be formed the compound of antibody and UCH-L1. The method also includes detecting resulting antibody-UCH-L1 compound.In the subject with traumatic brain injury, early stage is raw Substance markers object increases in about 0 to about 6 hour after suspicious lesion, then decreases or increases again.In some embodiments, UCH- The presence of L1 after to head injury about 0, about 0.5 hour, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours or Start to present in about 6 hours.Sample can be the biological sample (serum or blood sample) from subject.

In some embodiments, which comprises the first sample and the second optional sample are measured, to survey The level of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in amount or the first sample of detection and/or the second sample, wherein the first sample First time point of the product after to head suspicious lesion in 24 hours be derived from subject and the second sample first time point it The second time point afterwards is derived from subject in about 3 hours to about 6 hours such as after first time point；And by determining wound Property the severity of brain injury determines whether subject suffers from the damage to head.(i) when in the first sample and/or the second sample When UCH-L1 level is higher than the reference levels of UCH-L1, subject is determined as with moderate to severe trauma cerebral injury, and And subject is determined as suffering from when the UCH-L1 level in the first sample and/or the second sample is lower than the reference levels of UCH-L1 There is mild trauma cerebral injury；(ii) have when from the UCH-L1 level in the first sample to the UCH-L1 level in the second sample It is statistically significant that subject is determined as with moderate to severe trauma cerebral injury when increasing or decreasing, and when from UCH-L1 level in first sample to the UCH-L1 level in the second sample it is not statistically significant when increasing or decreasing, Subject is determined as with mild trauma cerebral injury；(iii) when the level of UCH-L1 subtracts from the first sample to the second sample When less or increasing at least absolute magnitude, subject is determined as with moderate to severe trauma cerebral injury, or when UCH-L1's When level does not decrease or increase at least absolute magnitude from the first sample to the second sample, subject is determined as with mild trauma Property cerebral injury；It (iv), will when the level of UCH-L1 decreases or increases at least the first absolute magnitude from the first sample to the second sample Subject is determined as with moderate to severe trauma cerebral injury, or horizontal from the first sample to the second sample as UCH-L1 When decreasing or increasing at least the second absolute magnitude, subject is determined as with mild trauma cerebral injury；Or (v) when in sample When UCH-L1 level is higher than the reference levels of UCH-L1 or when the UCH-L1 from the sample that first time point obtains is horizontal When dramatically increasing or reduce X amount to the UCH-L1 level in the sample that the second time point obtained, subject is determined as suffering from Moderate to severe trauma cerebral injury, and when the UCH-L1 level in sample is lower than the reference levels of UCH-L1 or when from UCH-L1 level in the sample that first time point obtains does not have to the UCH-L1 level in the sample that the second time point obtained When dramatically increasing or reduce greater than X amount, subject is determined as with mild trauma cerebral injury.In some embodiments In, the first absolute magnitude and the second absolute magnitude can be identical.In some embodiments, the first absolute magnitude and the second absolute magnitude It can be different.As used herein, " X " refers to the integer with non-zero value.As used herein, " determine whether subject suffers from Mild trauma cerebral injury or moderate are to severe trauma cerebral injury " refer to using the method (with or without other sides Method) determine that subject is more likely to slight TBI or moderate to severe TBI.

In some embodiments, subject received Golmud-Lhasa Pipeline before or after being measured and commented Point.In some embodiments, it is scored based on Golmud-Lhasa Pipeline, subject is doubtful with moderate to severe trauma brain Damage.In some embodiments, reference levels are associated with the subject of moderate to severe trauma cerebral injury.In In some embodiments, by analysis respective sample, using corresponding measuring method type identification reference levels come by reference levels with Subject with moderate to severe trauma cerebral injury is associated.

Optionally, in some embodiments, subject can not receive Glasgow dusk before being measured It is confused scale score.Optionally, in some embodiments, subject can not receive Glasgow after being measured Glasgow coma scale.In addition, in other embodiments, subject can receive Cranial Computed Tomography before being measured.In addition, In In other embodiment, subject can not receive Cranial Computed Tomography before being measured.Further, subject can be with Golmud-Lhasa Pipeline or Cranial Computed Tomography were not received before being measured.Still in further embodiment, tested Person can receive Golmud-Lhasa Pipeline before being measured but not receive Cranial Computed Tomography.Further implementing again In mode, subject can receive Cranial Computed Tomography before being measured but not receive Golmud-Lhasa Pipeline.More In further embodiment, subject can receive Cranial Computed Tomography and Golmud-Lhasa Pipeline before being measured.

In some embodiments, reference levels are associated with the scoring of the Golmud-Lhasa Pipeline of 3-12.Some It in embodiment, is scored based on Golmud-Lhasa Pipeline, subject is doubtful to suffer from mild trauma cerebral injury.In some implementations It is in mode, reference levels are associated with the subject of mild trauma cerebral injury.In some embodiments, it will refer to It is horizontal associated with the scoring of the Golmud-Lhasa Pipeline of 13-15.In some embodiments, by analyzing respective sample, making It is with corresponding measuring method type identification reference levels that reference levels are associated with the subject of mild trauma cerebral injury.

In general, reference levels are also used as the base of the result obtained when the UCH-L1 of assessment measurement test sample It is quasi-.In general, when carrying out this compare, reference levels obtain in the following manner: with enough numbers and in felicity condition Lower operation particular assay method so that can by the moment or terminal of analyte existence, amount or concentration and TBI or with spy Calibration will is associated or is associated with.In general, obtaining reference levels with the measurement to reference subject (or subject group). Measured UCH-L1 may include UCH-L1, its segment, its catabolite and/or its enzymatic lysis product.

In some embodiments, reference levels be by between at least about 85% to about 100% sensibility and What the measuring method of the specificity between at least about 30% to about 100% determined.In some embodiments, sensibility is at least about 85.0%, at least about 87.5%, at least about 90.0%, at least about 95.0%, at least about 99.0%, at least about 99.1%, at least About 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, extremely Few about 99.8%, at least about 99.9% or about 100.0%.In some embodiments, specificity is at least about 30.0%, at least About 31.0%, at least about 32.0%, at least about 33.0%, at least about 34.0%, at least about 35.0%, at least about 36.0%, extremely Few about 37.0%, at least about 38.0%, at least about 39.0%, at least about 40.0%, at least about 45.0%, at least about 50.0%, At least about 55.0%, at least about 60.0%, at least about 65.0%, at least about 70.0%, at least about 75.0%, at least about 80.0%, at least about 85.0%, at least about 90.0%, at least about 91.0%, at least about 92.0%, at least about 93.0%, at least About 94.0%, at least about 95.0%, at least about 96.0%, at least about 97.0%, at least about 98.0%, at least about 99.0%, extremely Few about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, At least about 99.7%, at least about 99.8%, at least about 99.9% or about 100.0%.For example, sensibility be at least about 99% and Specificity is at least about 75%, and sensibility is at least about 99% and specificity is at least about 99% or sensibility is about 100% And specificity is about 100%.In some embodiments, be at least about 99% and specificity at least about 75%.Again In other embodiment, sensibility is at least about 99% and specificity is at least about 99%.In still other embodiment, Sensibility is about 100% and specificity is about 100%.

In some embodiments, reference levels can be at least about 50pg/mL between about 12000pg/mL.Some In embodiment, reference levels can at least about 50pg/mL between about 12000pg/mL, at least about 50pg/mL to about Between 11000pg/mL, at least about 50pg/mL between about 10000pg/mL, at least about 50pg/mL to about 9500pg/mL Between, at least about 50pg/mL between about 9000pg/mL, at least about 50pg/mL between about 8500pg/mL, at least About 50pg/mL between about 8000pg/mL, at least about 50pg/mL between about 7500pg/mL, at least about 50pg/mL extremely Between about 7000pg/mL, at least about 50pg/mL between about 6500pg/mL, at least about 50pg/mL to about 6000pg/mL Between, at least about 50pg/mL between about 5500pg/mL, at least about 50pg/mL between about 5000pg/mL, at least About 50pg/mL between about 4500pg/mL, at least about 50pg/mL between about 4000pg/mL, at least about 50pg/mL extremely Between about 3500pg/mL, at least about 50pg/mL between about 3000pg/mL, at least about 50pg/mL to about 2500pg/mL Between, at least about 50pg/mL between about 2000pg/mL, at least about 50pg/mL between about 1500pg/mL, at least About 50pg/mL between about 1000pg/mL, at least about 50pg/mL between about 900pg/mL, at least about 50pg/mL extremely Between about 800pg/mL, at least about 50pg/mL between about 700pg/mL, at least about 50pg/mL to about 600pg/mL it Between, at least about 50pg/mL between about 500pg/mL, at least about 100pg/mL between about 12000pg/mL, at least About 100pg/mL between about 11000pg/mL, at least about 100pg/mL between about 10000pg/mL, at least about 100pg/mL between about 9500pg/mL, at least about 100pg/mL between about 9000pg/mL, at least about 100pg/mL To between about 8500pg/mL, at least about 100pg/mL between about 8000pg/mL, at least about 100pg/mL to about Between 7500pg/mL, at least about 100pg/mL between about 7000pg/mL, at least about 100pg/mL to about 6500pg/mL Between, at least about 100pg/mL between about 6000pg/mL, at least about 100pg/mL between about 5500pg/mL, extremely Few about 100pg/mL between about 5000pg/mL, at least about 100pg/mL between about 4500pg/mL, at least about 100pg/mL between about 4000pg/mL, at least about 100pg/mL between about 3500pg/mL, at least about 100pg/mL To between about 3000pg/mL, at least about 100pg/mL between about 2500pg/mL, at least about 100pg/mL to about Between 2000pg/mL, at least about 100pg/mL between about 1500pg/mL, at least about 100pg/mL to about 1000pg/mL Between, at least about 100pg/mL between about 900pg/mL, at least about 100pg/mL between about 800pg/mL, at least About 100pg/mL between about 700pg/mL, at least about 100pg/mL between about 600pg/mL, at least about 100pg/mL To between about 500pg/mL, at least about 150pg/mL between about 12000pg/mL, at least about 150pg/mL to about Between 11000pg/mL, at least about 150pg/mL between about 10000pg/mL, at least about 150pg/mL to about 9500pg/ Between mL, at least about 150pg/mL between about 9000pg/mL, at least about 150pg/mL between about 8500pg/mL, In At least about 150pg/mL between about 8000pg/mL, at least about 150pg/mL between about 7500pg/mL, at least about 150pg/mL between about 7000pg/mL, at least about 150pg/mL between about 6500pg/mL, at least about 150pg/mL To between about 6000pg/mL, at least about 150pg/mL between about 5500pg/mL, at least about 150pg/mL to about Between 5000pg/mL, at least about 150pg/mL between about 4500pg/mL, at least about 150pg/mL to about 4000pg/mL Between, at least about 150pg/mL between about 3500pg/mL, at least about 150pg/mL between about 3000pg/mL, extremely Few about 150pg/mL between about 2500pg/mL, at least about 150pg/mL between about 2000pg/mL, at least about 150pg/mL between about 1500pg/mL, at least about 150pg/mL between about 1000pg/mL, at least about 150pg/mL To between about 900pg/mL, at least about 150pg/mL between about 800pg/mL, at least about 150pg/mL to about 700pg/ Between mL, at least about 150pg/mL between about 600pg/mL, at least about 150pg/mL between about 500pg/mL, extremely Few about 200pg/mL between about 12000pg/mL, at least about 200pg/mL between about 11000pg/mL, at least about 200pg/mL between about 10000pg/mL, at least about 200pg/mL between about 9500pg/mL, at least about 200pg/mL To between about 9000pg/mL, at least about 200pg/mL between about 8500pg/mL, at least about 200pg/mL to about Between 8000pg/mL, at least about 200pg/mL between about 7500pg/mL, at least about 200pg/mL to about 7000pg/mL Between, at least about 200pg/mL between about 6500pg/mL, at least about 200pg/mL between about 6000pg/mL, extremely Few about 200pg/mL between about 5500pg/mL, at least about 200pg/mL between about 5000pg/mL, at least about 200pg/mL between about 4500pg/mL, at least about 200pg/mL between about 4000pg/mL, at least about 200pg/mL To between about 3500pg/mL, at least about 200pg/mL between about 3000pg/mL, at least about 200pg/mL to about Between 2500pg/mL, at least about 200pg/mL between about 2000pg/mL, at least about 200pg/mL to about 1500pg/mL Between, at least about 200pg/mL between about 1000pg/mL, at least about 200pg/mL between about 900pg/mL, extremely Few about 200pg/mL between about 800pg/mL, at least about 200pg/mL between about 700pg/mL, at least about 200pg/ ML between about 600pg/mL, at least about 200pg/mL between about 500pg/mL, at least about 300pg/mL to about Between 12000pg/mL, at least about 300pg/mL between about 11000pg/mL, at least about 300pg/mL to about Between 10000pg/mL, at least about 300pg/mL between about 9500pg/mL, at least about 300pg/mL to about 9000pg/ Between mL, at least about 300pg/mL between about 8500pg/mL, at least about 300pg/mL between about 8000pg/mL, In At least about 300pg/mL between about 7500pg/mL, at least about 300pg/mL between about 7000pg/mL, at least about 300pg/mL between about 6500pg/mL, at least about 300pg/mL between about 6000pg/mL, at least about 300pg/mL To between about 5500pg/mL, at least about 300pg/mL between about 5000pg/mL, at least about 300pg/mL to about Between 4500pg/mL, at least about 300pg/mL between about 4000pg/mL, at least about 300pg/mL to about 3500pg/mL Between, at least about 300pg/mL between about 3000pg/mL, at least about 300pg/mL between about 2500pg/mL, extremely Few about 300pg/mL between about 2000pg/mL, at least about 300pg/mL between about 1500pg/mL, at least about 300pg/mL between about 1000pg/mL, at least about 300pg/mL between about 900pg/mL, at least about 300pg/mL extremely Between about 800pg/mL, at least about 300pg/mL between about 700pg/mL, at least about 300pg/mL to about 600pg/mL Between, at least about 300pg/mL between about 500pg/mL, at least about 400pg/mL between about 12000pg/mL, extremely Few about 400pg/mL between about 11000pg/mL, at least about 400pg/mL between about 10000pg/mL, at least about 400pg/mL between about 9500pg/mL, at least about 400pg/mL between about 9000pg/mL, at least about 400pg/mL To between about 8500pg/mL, at least about 400pg/mL between about 8000pg/mL, at least about 400pg/mL to about Between 7500pg/mL, at least about 400pg/mL between about 7000pg/mL, at least about 400pg/mL to about 6500pg/mL Between, at least about 400pg/mL between about 6000pg/mL, at least about 400pg/mL between about 5500pg/mL, extremely Few about 400pg/mL between about 5000pg/mL, at least about 400pg/mL between about 4500pg/mL, at least about 400pg/mL between about 4000pg/mL, at least about 400pg/mL between about 3500pg/mL, at least about 400pg/mL To between about 3000pg/mL, at least about 400pg/mL between about 2500pg/mL, at least about 400pg/mL to about Between 2000pg/mL, at least about 400pg/mL between about 1500pg/mL, at least about 400pg/mL to about 1000pg/mL Between, at least about 400pg/mL between about 900pg/mL, at least about 400pg/mL between about 800pg/mL, at least About 400pg/mL between about 700pg/mL, at least about 400pg/mL between about 600pg/mL, at least about 400pg/mL To between about 500pg/mL, at least about 500pg/mL between about 12000pg/mL, at least about 500pg/mL to about Between 11000pg/mL, at least about 500pg/mL between about 10000pg/mL, at least about 500pg/mL to about 9500pg/ Between mL, at least about 500pg/mL between about 9000pg/mL, at least about 500pg/mL between about 8500pg/mL, In At least about 500pg/mL between about 8000pg/mL, at least about 500pg/mL between about 7500pg/mL, at least about 500pg/mL between about 7000pg/mL, at least about 500pg/mL between about 6500pg/mL, at least about 500pg/mL To between about 6000pg/mL, at least about 500pg/mL between about 5500pg/mL, at least about 500pg/mL to about Between 5000pg/mL, at least about 500pg/mL between about 4500pg/mL, at least about 500pg/mL to about 4000pg/mL Between, at least about 500pg/mL between about 3500pg/mL, at least about 500pg/mL between about 3000pg/mL, extremely Few about 500pg/mL between about 2500pg/mL, at least about 500pg/mL between about 2000pg/mL, at least about 500pg/mL between about 1500pg/mL, at least about 500pg/mL between about 1000pg/mL, at least about 500pg/mL To between about 900pg/mL, at least about 500pg/mL between about 800pg/mL, at least about 500pg/mL to about 700pg/ Between mL, at least about 500pg/mL between about 600pg/mL, at least about 1000pg/mL between about 12000pg/mL, At least about 1000pg/mL between about 11000pg/mL, at least about 1000pg/mL between about 10000pg/mL, extremely Few about 1000pg/mL between about 9500pg/mL, at least about 1000pg/mL between about 9000pg/mL, at least about 1000pg/mL between about 8500pg/mL, at least about 1000pg/mL between about 8000pg/mL, at least about 1000pg/ ML between about 7500pg/mL, at least about 1000pg/mL between about 7000pg/mL, at least about 1000pg/mL to about Between 6500pg/mL, at least about 1000pg/mL between about 6000pg/mL, at least about 1000pg/mL to about 5500pg/ Between mL, at least about 1000pg/mL between about 5000pg/mL, at least about 1000pg/mL between about 4500pg/mL, At least about 1000pg/mL between about 4000pg/mL, at least about 1000pg/mL between about 3500pg/mL, at least About 1000pg/mL between about 3000pg/mL, at least about 1000pg/mL between about 2500pg/mL, at least about 1000pg/mL between about 2000pg/mL, at least about 1000pg/mL between about 1500pg/mL, at least about 2000pg/ ML between about 12000pg/mL, at least about 2000pg/mL between about 11000pg/mL, at least about 2000pg/mL extremely Between about 10000pg/mL, at least about 2000pg/mL between about 9500pg/mL, at least about 2000pg/mL to about Between 9000pg/mL, at least about 2000pg/mL between about 8500pg/mL, at least about 2000pg/mL to about 8000pg/ Between mL, at least about 2000pg/mL between about 7500pg/mL, at least about 2000pg/mL between about 7000pg/mL, At least about 2000pg/mL between about 6500pg/mL, at least about 2000pg/mL between about 6000pg/mL, at least About 2000pg/mL between about 5500pg/mL, at least about 2000pg/mL between about 5000pg/mL, at least about 2000pg/mL between about 4500pg/mL, at least about 2000pg/mL between about 4000pg/mL, at least about 2000pg/ ML between about 3500pg/mL, at least about 2000pg/mL between about 3000pg/mL or at least about 2000pg/mL to about Between 2500pg/mL.For example, reference levels can be at least about 65pg/mL between about 9019pg/mL.In some embodiment party In formula, reference levels can be at least about 50pg/mL, at least about 55pg/mL, at least about 60pg/mL, at least about 65pg/mL, extremely Few about 70pg/mL, at least about 75pg/mL, at least about 80pg/mL, at least about 85pg/mL, at least about 90pg/mL, at least about 95pg/mL, at least about 96pg/mL, at least about 97pg/mL, at least about 98pg/mL, at least about 99pg/mL, at least about 100pg/ ML, at least about 150pg/mL, at least about 200pg/mL, at least about 205pg/mL, at least about 206pg/mL, at least about 207pg/ ML, at least about 208pg/mL, at least about 209pg/mL, at least about 210pg/mL, at least about 220pg/mL, at least about 230pg/ ML, at least about 238pg/mL, at least about 240pg/mL, at least about 250pg/mL, at least about 300pg/mL, at least about 311pg/ ML, at least about 320pg/mL, at least about 330pg/mL, at least about 350pg/mL, at least about 360pg/mL, at least about 370pg/ ML, at least about 380pg/mL, at least about 390pg/mL, at least about 400pg/mL, at least about 450pg/mL, at least about 500pg/ ML, at least about 505pg/mL, at least about 506pg/mL, at least about 507pg/mL, at least about 508pg/mL, at least about 509pg/ ML, at least about 510pg/mL, at least about 520pg/mL, at least about 530pg/mL, at least about 540pg/mL, at least about 550pg/ ML, at least about 560pg/mL, at least about 565pg/mL, at least about 569pg/mL, at least about 570pg/mL, at least about 700pg/ ML, at least about 710pg/mL, at least about 720pg/mL, at least about 730pg/mL, at least about 740pg/mL, at least about 750pg/ ML, at least about 800pg/mL, at least about 850pg/mL, at least about 900pg/mL, at least about 950pg/mL, at least about 1000pg/ ML, at least about 1100pg/mL, at least about 1200pg/mL, at least about 1300pg/mL, at least about 1400pg/mL, at least about 1500pg/mL, at least about 1600pg/mL, at least about 1700pg/mL, at least about 1800pg/mL, at least about 1900pg/mL, extremely Few about 2000pg/mL, at least about 2500pg/mL, at least about 3000pg/mL, at least about 3500pg/mL, at least about 4000pg/ ML, at least about 4500pg/mL, at least about 5000pg/mL, at least about 5500pg/mL, at least about 6000pg/mL, at least about 6500pg/mL, at least about 7000pg/mL, at least about 7500pg/mL, at least about 8000pg/mL, at least about 8500pg/mL, extremely Few about 9000pg/mL, at least about 9010pg/mL, at least about 9020pg/mL, at least about 9500pg/mL, at least about 10000pg/ ML, at least about 11000pg/mL or at least about 12000pg/mL.In some embodiments, reference levels are at least about 50pg/ ML is between about 12000pg/mL.In other embodiments, reference levels at least about 65pg/mL to about 9019pg/mL it Between.

In some embodiments, the first sample obtains in 24 hours after suspicious lesion, and the second sample is first It is obtained in about 3 to about 6 hours after sample and reference levels is by with certain sensitivity as described above and specificity What measuring method determined.For example, the first sample can after suspicious lesion in about 0 hour, in about 30 minutes, in about 1 hour, about In 2 hours, in about 3 hours, in about 4 hours, in about 5 hours, in about 6 hours, in about 7 hours, in about 8 hours, about 9 hours It is interior, in about 10 hours, in about 11 hours, in about 12 hours, in about 13 hours, in about 14 hours, in about 15 hours, about 16 hours It is interior, in about 17 hours, in about 18 hours, in about 19 hours, in about 20 hours, in about 21 hours, in about 22 hours, about 23 hours It is interior, obtain in about 24 hours or more than about 24 hours.

In some embodiments, the first sample after suspicious lesion in about 0 to about 1 hour, in about 0 to about 2 hour, about In 0 to about 3 hour, in about 0 to about 4 hour, in about 0 to about 5 hour, in about 0 to about 6 hour, in about 0 to about 7 hour, about 0 In to about 8 hours, in about 0 to about 9 hour, in about 0 to about 10 hour, in about 0 to about 11 hour, in about 0 to about 12 hour, about It is taken in 0 to about 18 hour, in about 6 to about 12 hours, in about 12 to about 18 hours, in about 18 to about 24 hours or greater than 24 hours It obtains and reference levels is by the sensibility with certain sensitivity and specificity, such as between at least about 85% and 100% Specificity, such as about 100% special sensibility and at least about 75% specificity between at least about 30% and 100%, extremely The measurement of few about 99% sensibility and at least about 75% specificity or about 100% sensibility and about 100% specificity What method determined.

In some embodiments, the first sample can obtain in about 0 to about 6 hour, and reference levels can be about 311pg/mL and measuring method have about 100% sensibility and at least about 33% specificity.In some embodiments, A sample can obtain in about 0 to about 6 hour, and reference levels can be about 509pg/mL and measuring method has about 100% Sensibility and at least about 66% specificity.In some embodiments, the first sample can obtain in about 0 to about 6 hour , reference levels can be about 710pg/mL and measuring method has about 100% sensibility and at least about 92% specificity. In some embodiments, the first sample can obtain in about 0 to about 6 hour, and reference levels can be about 9019pg/mL simultaneously And measuring method has about 100% sensibility and about 100% specificity.

In some embodiments, the first sample can be obtained between after suspicious lesion about 6 hours to 12 hours and Reference levels are between about 65pg/mL and about 9019pg/mL and are quick between at least about 85% to about 100% by having What the measuring method of the specificity between perception and at least about 30% and about 100% determined.In some embodiments, the first sample It is obtained between after suspicious lesion about 6 hours to 12 hours and reference levels is about 98pg/mL and is by having about 100% sensibility and at least about 30% specificity measuring method determine.In some embodiments, the first sample is being doubted It is about 209pg/mL like acquirement and reference levels between after damage about 6 hours to 12 hours and is by having about 100% Sensibility and at least about 63% specificity measuring method determine.In some embodiments, the first sample is in doubtful damage It is obtained between about 6 hours to 12 hours after wound and reference levels is about 238pg/mL and is by having at least about 90% Sensibility and at least about 70% specificity measuring method determine.In some embodiments, the first sample is in suspicious lesion Afterwards between about 6 hours to 12 hours obtain and reference levels be about 569pg/mL and be by at least about 90% it is quick What the measuring method of perception and at least about 96% specificity determined.

In some embodiments, the first sample is obtained in first time point and obtain the second sample at the second time point To determine whether subject suffers from slight or moderate to severe TBI.In some embodiments, first time point is damaged to head About 0 to about 24 hour after wound or suspicious lesion.For example, first time point can be after suspicious lesion it is about 0 to about 24 small When, about 0 to about 20 hour, about 0 to about 18 hour, about 0 to about 16 hour, about 0 to about 14 hour, about 0 to about 12 hour, about 0 To about 10 hours, about 0 to about 8 hour, about 0 to about 6 hour, about 0 to about 4 hour, about 0 to about 2 hour, about 0 to about 1 hour, About 0 to about 1.5 hour, about 0.5 hour to about 24 hours, about 0.5 hour to about 20 hours, about 0.5 hour to about 18 hours, about 0.5 hour to about 16 hours, about 0.5 hour to about 14 hours, about 0.5 hour to about 12 hours, it is about 0.5 hour to about 10 small When, about 0.5 hour to about 8 hours, about 0.5 hour to about 6 hours, about 0.5 hour to about 4 hours, it is about 0.5 hour to about 2 small When, about 0.5 hour to about 1 hour, about 0.5 hour to about 1.5 hours, about 1 hour to about 24 hours, it is about 1 hour to about 20 small When, about 1 hour to about 18 hours, about 1 hour to about 16 hours, about 1 hour to about 14 hours, about 1 hour to about 12 hours, about 1 hour to about 10 hours, about 1 hour to about 8 hours, about 1 hour to about 6 hours, about 1 hour to about 4 hours, about 1 hour extremely About 2 hours, about 2 hours to about 24 hours, about 2 hours to about 20 hours, about 2 hours to about 18 hours, it is about 2 hours to about 16 small When, about 2 hours to about 14 hours, about 2 hours to about 12 hours, about 2 hours to about 10 hours, about 2 hours to about 8 hours, about 2 Hour to about 6 hours, about 2 hours to about 4 hours, about 3 hours to about 24 hours, about 3 hours to about 20 hours, about 3 hours extremely About 18 hours, about 3 hours to about 16 hours, about 3 hours to about 14 hours, about 3 hours to about 12 hours, about 3 hours to about 10 Hour, about 3 hours to about 8 hours, about 3 hours to about 6 hours, about 3 hours to about 4 hours, about 4 hours to about 24 hours, about 4 Hour was to about 20 hours, about 4 hours to about 18 hours, about 4 hours to about 16 hours, about 4 hours to about 14 hours, about 4 hours To about 12 hours, about 4 hours to about 10 hours, about 4 hours to about 8 hours, about 4 hours to about 6 hours, about 6 hours to about 24 Hour, about 6 hours to about 20 hours, about 6 hours to about 18 hours, about 6 hours to about 16 hours, about 6 hours to about 14 hours, About 6 hours to about 12 hours, about 6 hours to about 10 hours, about 6 hours to about 8 hours, about 12 hours to about 24 hours, about 12 Hour was to about 20 hours, about 12 hours to about 18 hours, about 12 hours to about 16 hours, about 12 hours to about 14 hours, about 18 Between hour to about 24 hours or it is greater than 24 hours.For example, first time point can be between 0 hour to 6 hours, about 6 small Between 12 hours, between about 12 hours to 18 hours or between about 18 hours to 24 hours.In some embodiments, First time point is between about 0 hour to about 6 hours.In some embodiments, first time point is small at about 6 hours to about 12 When between.In some embodiments, first time point is between about 12 hours to about 18 hours.In other embodiments, First time point is between about 18 hours to about 24 hours.

In some embodiments, the second time point was about 1 hour to about 10 hours after first time point, such as first About 3 hours to about 6 hours after time point.In some embodiments, the second time point was about 1 small after first time point When, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours or about 10 hours.

In some embodiments, from the UCH-L1 level in the first sample that first time point obtains at second Between to put statistically significant the increasing or decreasing of the UCH-L1 level in the second sample of acquirement be at least about 0.1 times, at least About 0.2 times, at least about 0.3 times, at least about 0.4 times, at least about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, at least about 1 times, at least 1.5 times, at least about 2 times, at least About 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 Times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times or At least 20 times.In some embodiments, the UCH-L1 level from the first sample that first time point obtains is to second Horizontal statistically significant of the UCH-L1 in the second sample that time point obtains be reduced by least about 0.1 times, at least about 0.2 times, At least about 0.3 times, at least about 0.4 times, at least about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least About 0.73 times, at least about 0.8 times, at least about 0.9 times, at least about 1 times, at least 1.5 times, at least about 2 times, at least about 3 times, at least About 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times or at least 20 times Show subject with moderate to severe trauma cerebral injury.In some embodiments, from obtained in first time point UCH-L1 level in a sample is to horizontal statistically significant of the UCH-L1 in the second sample that the second time point obtained At least about 0.1 times, at least about 0.2 times, at least about 0.3 times, at least about 0.4 times, at least about 0.5 times, at least about 0.55 times of increase, At least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, at least about 1 times, at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least 8 times, extremely Few 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 Again, show subject with moderate to severe trauma cerebral injury at least 18 times, at least 19 times or at least 20 times.

In some embodiments, from the UCH-L1 level in the first sample that first time point obtains at second Between put statistically significant the increasing or decreasing of the UCH-L1 level in the second sample of acquirement and be less than about 0.1 times, be less than About 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, less than about 1 times, less than 1.5 times, less than about 2 times, be less than About 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 times, be less than about 7 times, less than 8 times, less than 9 times, less than 10 times, less than 11 Times, less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 times, less than 18 times, less than 19 times or Less than 20 times.In some embodiments, the UCH-L1 level from the first sample that first time point obtains is to second Time point obtain the second sample in UCH-L1 level statistically significant reduction be less than about 0.1 times, less than about 0.2 times, Less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least About 0.73 times, at least about 0.8 times, at least about 0.9 times, less than about 1 times, less than 1.5 times, less than about 2 times, less than about 3 times, be less than About 4 times, less than about 5 times, less than about 6 times, less than about 7 times, less than about 8 times, less than 9 times, less than 10 times, less than 11 times, be less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 times, less than 18 times, less than 19 times or less than 20 times Show subject with moderate to severe trauma cerebral injury.In some embodiments, from obtained in first time point UCH-L1 level in a sample is to horizontal statistically significant of the UCH-L1 in the second sample that the second time point obtained Increase be less than about 0.1 times, be less than about 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, At least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, less than about 1 times, be less than about 1.5 times, be less than about 2 times, be less than about 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 times, be less than about 7 times, less than 8 times, it is small In 9 times, less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 Again, show subject with moderate to severe trauma cerebral injury less than 18 times, less than 19 times or less than 20 times.In some implementations In mode, from the UCH-L1 level in the first sample that first time point obtains to the second sample obtained at the second time point In not statistically significant the increasing or decreasing of UCH-L1 level show subject with mild trauma cerebral injury.

In some embodiments, from the UCH-L1 level in the second sample that the second time point obtained at first Between put the UCH-L1 level in the first sample of acquirement statistically significant increasing or decreasing be more than about 0.1 times, be more than About 0.2 times, more than about 0.3 times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, more than about 1 times, more than 1.5 times, more than about 2 times, be more than About 3 times, more than about 4 times, more than about 5 times, more than about 6 times, more than about 7 times, more than 8 times, more than 9 times, more than 10 times, more than 11 Times, more than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 times, more than 18 times, more than 19 times or More than 20 times.In some embodiments, the UCH-L1 level from the second sample that the second time point obtained is to first Horizontal statistically significant of the UCH-L1 in the first sample that time point obtains reduce by more than about 0.1 times, more than about 0.2 times, More than about 0.3 times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least About 0.73 times, at least about 0.8 times, at least about 0.9 times, more than about 1 times, more than 1.5 times, more than about 2 times, more than about 3 times, be more than About 4 times, more than about 5 times, more than about 6 times, more than about 7 times, more than about 8 times, more than 9 times, more than 10 times, more than 11 times, be more than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 times, more than 18 times, more than 19 times or more than 20 times Show subject with moderate to severe trauma cerebral injury.In some embodiments, from obtained at the second time point UCH-L1 level in two samples is to horizontal statistically significant of the UCH-L1 in the first sample that first time point obtains Increase above about 0.1 times, more than about 0.2 times, more than about 0.3 times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, At least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, more than about 1 times, be more than about 1.5 times, more than about 2 times, more than about 3 times, more than about 4 times, more than about 5 times, more than about 6 times, more than about 7 times, be more than 8 times, surpass Cross 9 times, more than 10 times, more than 11 times, more than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 Again, show subject with moderate to severe trauma cerebral injury more than 18 times, more than 19 times or more than 20 times.In some realities It applies in mode, from the UCH-L1 level in the second sample that the second time point obtained to the first sample obtained in first time point Not statistically significant the increasing or decreasing of UCH-L1 level in product shows subject with mild trauma cerebral injury.

In some embodiments, first time point is about 0 to about 12 hour after suspicious lesion and statistically aobvious The increase of work is from the second time point to first time point more than about 1 times.In some embodiments, first time point is doubtful About 0 to about 12 hour after damage and statistically significant increase are more than about from the second time point to first time point 0.73 times.In some embodiments, first time point is about 0 to about 12 hour after suspicious lesion, the reference water of UCH-L1 It puts down between about 350pg/mL and about 550pg/mL, and statistically significant increase is from the second time point at the first time Point is more than about 0.73 times.

In some embodiments, absolute magnitude can be by having the sensibility between at least about 70% to about 100% What the measuring method of the specificity between at least about 30% to about 100% determined.In some embodiments, sensibility is at least About 70.0%, at least about 75.0%, at least about 80.0%, at least about 85.0%, at least about 87.5%, at least about 90.0%, extremely Few about 95.0%, at least about 99.0%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, At least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9% or at least about 100.0%.In some embodiments, specificity at least about 30.0%, at least about 31.0%, at least about 32.0%, at least About 33.0%, at least about 34.0%, at least about 35.0%, at least about 36.0%, at least about 37.0%, at least about 38.0%, extremely Few about 39.0%, at least about 40.0%, at least about 45.0%, at least about 50.0%, at least about 55.0%, at least about 60.0%, At least about 65.0%, at least about 70.0%, at least about 75.0%, at least about 80.0%, at least about 85.0%, at least about 90.0%, at least about 91.0%, at least about 92.0%, at least about 93.0%, at least about 94.0%, at least about 95.0%, at least About 96.0%, at least about 97.0%, at least about 98.0%, at least about 99.0%, at least about 99.1%, at least about 99.2%, extremely Few about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, At least about 99.9% or at least about 100.0%.For example, sensibility is at least about 99% and specificity is at least about 75%, it is quick Perception is at least about 99% and specificity is at least about 99% or sensibility is about 100% and specificity is about 100%.

In some embodiments, absolute magnitude can be at least about 20pg/mL between about 6100pg/mL.In some realities Apply in mode, absolute magnitude can at least about 20pg/mL between about 6100pg/mL, at least about 20pg/mL to about Between 6000pg/mL, at least about 20pg/mL between about 5500pg/mL, at least about 20pg/mL to about 5000pg/mL it Between, at least about 20pg/mL between about 4500pg/mL, at least about 20pg/mL between about 4000pg/mL, at least about 20pg/mL between about 3500pg/mL, at least about 20pg/mL between about 3000pg/mL, at least about 20pg/mL to about Between 2500pg/mL, at least about 20pg/mL between about 2000pg/mL, at least about 20pg/mL to about 1500pg/mL it Between, at least about 20pg/mL between about 1000pg/mL, at least about 20pg/mL between about 900pg/mL, at least about 20pg/mL between about 800pg/mL, at least about 20pg/mL between about 700pg/mL, at least about 20pg/mL to about Between 600pg/mL, at least about 20pg/mL between about 500pg/mL, at least about 20pg/mL between about 400pg/mL, At least about 20pg/mL between about 300pg/mL, at least about 20pg/mL between about 200pg/mL, at least about 20pg/ ML between about 100pg/mL, at least about 20pg/mL between about 50pg/mL, at least about 25pg/mL to about 6100pg/ Between mL, at least about 25pg/mL between about 6000pg/mL, at least about 25pg/mL between about 5500pg/mL, extremely Few about 25pg/mL between about 5000pg/mL, at least about 25pg/mL between about 4500pg/mL, at least about 25pg/mL To between about 4000pg/mL, at least about 25pg/mL between about 3500pg/mL, at least about 25pg/mL to about 3000pg/ Between mL, at least about 25pg/mL between about 2500pg/mL, at least about 25pg/mL between about 2000pg/mL, extremely Few about 25pg/mL between about 1500pg/mL, at least about 25pg/mL between about 1000pg/mL, at least about 25pg/mL To between about 900pg/mL, at least about 25pg/mL between about 800pg/mL, at least about 25pg/mL to about 700pg/mL Between, at least about 25pg/mL between about 600pg/mL, at least about 25pg/mL between about 500pg/mL, at least about 25pg/mL between about 400pg/mL, at least about 25pg/mL between about 300pg/mL, at least about 25pg/mL to about Between 200pg/mL, at least about 25pg/mL between about 100pg/mL, at least about 25pg/mL between about 50pg/mL, At least about 50pg/mL between about 6100pg/mL, at least about 50pg/mL between about 6000pg/mL, at least about 50pg/mL between about 5500pg/mL, at least about 50pg/mL between about 5000pg/mL, at least about 50pg/mL to about Between 4500pg/mL, at least about 50pg/mL between about 4000pg/mL, at least about 50pg/mL to about 3500pg/mL it Between, at least about 50pg/mL between about 3000pg/mL, at least about 50pg/mL between about 2500pg/mL, at least about 50pg/mL between about 2000pg/mL, at least about 50pg/mL between about 1500pg/mL, at least about 50pg/mL to about Between 1000pg/mL, at least about 50pg/mL between about 900pg/mL, at least about 50pg/mL to about 800pg/mL it Between, at least about 50pg/mL between about 700pg/mL, at least about 50pg/mL between about 600pg/mL, at least about 50pg/mL between about 500pg/mL, at least about 50pg/mL between about 400pg/mL, at least about 50pg/mL to about Between 300pg/mL, at least about 50pg/mL between about 200pg/mL, at least about 50pg/mL between about 100pg/mL, At least about 100pg/mL between about 6100pg/mL, at least about 100pg/mL between about 6000pg/mL, at least about 100pg/mL between about 5500pg/mL, at least about 100pg/mL between about 5000pg/mL, at least about 100pg/mL To between about 4500pg/mL, at least about 100pg/mL between about 4000pg/mL, at least about 100pg/mL to about Between 3500pg/mL, at least about 100pg/mL between about 3000pg/mL, at least about 100pg/mL to about 2500pg/mL Between, at least about 100pg/mL between about 2000pg/mL, at least about 100pg/mL between about 1500pg/mL, extremely Few about 100pg/mL between about 1000pg/mL, at least about 100pg/mL between about 900pg/mL, at least about 100pg/ ML between about 800pg/mL, at least about 100pg/mL between about 700pg/mL, at least about 100pg/mL to about Between 600pg/mL, at least about 100pg/mL between about 500pg/mL, at least about 100pg/mL to about 400pg/mL it Between, at least about 100pg/mL between about 300pg/mL, at least about 100pg/mL between about 200pg/mL, at least about 129pg/mL between about 6100pg/mL, at least about 129pg/mL between about 6000pg/mL, at least about 129pg/mL To between about 5500pg/mL, at least about 129pg/mL between about 5000pg/mL, at least about 129pg/mL to about Between 4500pg/mL, at least about 129pg/mL between about 4000pg/mL, at least about 129pg/mL to about 3500pg/mL Between, at least about 129pg/mL between about 3000pg/mL, at least about 129pg/mL between about 2500pg/mL, extremely Few about 129pg/mL between about 2000pg/mL, at least about 129pg/mL between about 1500pg/mL, at least about 129pg/mL between about 1000pg/mL, at least about 129pg/mL between about 900pg/mL, at least about 129pg/mL extremely Between about 800pg/mL, at least about 129pg/mL between about 700pg/mL, at least about 129pg/mL to about 600pg/mL Between, at least about 129pg/mL between about 500pg/mL, at least about 129pg/mL between about 400pg/mL, at least About 129pg/mL between about 300pg/mL, at least about 129pg/mL between about 200pg/mL, at least about 200pg/mL To between about 6100pg/mL, at least about 200pg/mL between about 6000pg/mL, at least about 200pg/mL to about Between 5500pg/mL, at least about 200pg/mL between about 5000pg/mL, at least about 200pg/mL to about 4500pg/mL Between, at least about 200pg/mL between about 4000pg/mL, at least about 200pg/mL between about 3500pg/mL, extremely Few about 200pg/mL between about 3000pg/mL, at least about 200pg/mL between about 2500pg/mL, at least about 200pg/mL between about 2000pg/mL, at least about 200pg/mL between about 1500pg/mL, at least about 200pg/mL To between about 1000pg/mL, at least about 200pg/mL between about 900pg/mL, at least about 200pg/mL to about 800pg/ Between mL, at least about 200pg/mL between about 700pg/mL, at least about 200pg/mL between about 600pg/mL, extremely Few about 200pg/mL between about 500pg/mL, at least about 200pg/mL between about 400pg/mL, at least about 200pg/ ML between about 300pg/mL, at least about 300pg/mL between about 6100pg/mL, at least about 300pg/mL to about Between 6000pg/mL, at least about 300pg/mL between about 5500pg/mL, at least about 300pg/mL to about 5000pg/mL Between, at least about 300pg/mL between about 4500pg/mL, at least about 300pg/mL between about 4000pg/mL, extremely Few about 300pg/mL between about 3500pg/mL, at least about 300pg/mL between about 3000pg/mL, at least about 300pg/mL between about 2500pg/mL, at least about 300pg/mL between about 2000pg/mL, at least about 300pg/mL To between about 1500pg/mL, at least about 300pg/mL between about 1000pg/mL, at least about 300pg/mL to about Between 900pg/mL, at least about 300pg/mL between about 800pg/mL, at least about 300pg/mL to about 700pg/mL it Between, at least about 300pg/mL between about 600pg/mL, at least about 300pg/mL between about 500pg/mL, at least about 300pg/mL between about 400pg/mL, at least about 400pg/mL between about 6100pg/mL, at least about 400pg/mL extremely Between about 6000pg/mL, at least about 400pg/mL between about 5500pg/mL, at least about 400pg/mL to about 5000pg/ Between mL, at least about 400pg/mL between about 4500pg/mL, at least about 400pg/mL between about 4000pg/mL, In At least about 400pg/mL between about 3500pg/mL, at least about 400pg/mL between about 3000pg/mL, at least about 400pg/mL between about 2500pg/mL, at least about 400pg/mL between about 2000pg/mL, at least about 400pg/mL To between about 1500pg/mL, at least about 400pg/mL between about 1000pg/mL, at least about 400pg/mL to about Between 900pg/mL, at least about 400pg/mL between about 800pg/mL, at least about 400pg/mL to about 700pg/mL it Between, at least about 400pg/mL between about 600pg/mL, at least about 400pg/mL between about 500pg/mL, at least about 500pg/mL between about 6100pg/mL, at least about 500pg/mL between about 6000pg/mL, at least about 500pg/mL To between about 5500pg/mL, at least about 500pg/mL between about 5000pg/mL, at least about 500pg/mL to about Between 4500pg/mL, at least about 500pg/mL between about 4000pg/mL, at least about 500pg/mL to about 3500pg/mL Between, at least about 500pg/mL between about 3000pg/mL, at least about 500pg/mL between about 2500pg/mL, extremely Few about 500pg/mL between about 2000pg/mL, at least about 500pg/mL between about 1500pg/mL, at least about 500pg/mL between about 1000pg/mL, at least about 500pg/mL between about 900pg/mL, at least about 500pg/mL extremely To between about 700pg/mL or at least about 500pg/mL to about 600pg/mL between about 800pg/mL, at least about 500pg/mL Between.In some embodiments, absolute magnitude can be at least about 20pg/mL, at least about 21pg/mL, at least about 22pg/mL, At least about 23pg/mL, at least about 24pg/mL, at least about 25pg/mL, at least about 26pg/mL, at least about 27pg/mL, at least about 28pg/mL, at least about 29pg/mL, at least about 30pg/mL, at least about 35pg/mL, at least about 40pg/mL, at least about 45pg/ ML, at least about 50pg/mL, at least about 55pg/mL, at least about 60pg/mL, at least about 65pg/mL, at least about 70pg/mL, at least About 75pg/mL, at least about 80pg/mL, at least about 85pg/mL, at least about 90pg/mL, at least about 95pg/mL, at least about 100pg/mL, at least about 110pg/mL, at least about 120pg/mL, at least about 129pg/mL, at least about 130pg/mL, at least about 140pg/mL, at least about 150pg/mL, at least about 200pg/mL, at least about 250pg/mL, at least about 300pg/mL, at least about 350pg/mL, at least about 400pg/mL, at least about 450pg/mL, at least about 500pg/mL, at least about 550pg/mL, at least about 600pg/mL, at least about 700pg/mL, at least about 800pg/mL, at least about 900pg/mL, at least about 1000pg/mL, at least about 1500pg/mL, at least about 2000pg/mL, at least about 2500pg/mL, at least about 2528pg/mL, at least about 3000pg/mL, extremely Few about 4000pg/mL, at least about 5000pg/mL, at least about 6000pg/mL or at least about 6100pg/mL.

In some embodiments, the first sample can obtain in about 0 to about 6 hour, and reference levels can be Pass through the about 2528pg/mL that there is the measuring method of about 100% sensibility and about 100% specificity to determine.In some implementations In mode, the first sample can obtain in about 0 to about 6 hour, and reference levels can be by having at least about 70% Sensibility and at least about 92% specificity measuring method determine about 129pg/mL.In some embodiments, the first sample Product can obtain in about 0 to about 10 hour, and reference levels can be through the sensibility and at least about with about 100% The about 25pg/mL that the measuring method of 36% specificity determines.In some embodiments, the first sample can be about 0 to about 11 It is obtained in hour, and reference levels can be the survey by having about 100% sensibility and at least about 32% specificity Determine the about 25pg/mL that method determines.In some embodiments, the first sample can obtain in about 0 to about 12 hour, and join Examine the horizontal pact that can be by having the measuring method of at least about 75% sensibility and at least about 76% specificity to determine 129pg/mL。

In some embodiments, the method further includes being assessed as suffering from the treatment of traumatic brain injury therapy Moderate to severe trauma cerebral injury human experimenter, as described below.In some embodiments, the method is further wrapped It includes monitoring and is assessed as the human experimenter with mild trauma cerebral injury, as described below.

The property of the measuring method used in methods described herein is not critical, and test can be it is as known in the art Any measuring method, such as immunoassay, clinical chemistry measuring method, Single Molecule Detection measuring method, the Western Immuno precipitation method, Immunoelectrophoresis, chemical analysis, SDS-PAGE and western blot analysis or Western Immuno decoration method, electrophoretic analysis Method, protein determination, competitive binding assay, functional protein measuring method or chromatography or spectroscopic methodology, such as efficient liquid Phase chromatography (HPLC) or liquid chromatography-mass spectrography (LC/MS).In addition, measuring method can be used in the form of clinical chemistry, such as It will be by known to persons of ordinary skill in the art.Such measuring method is described in further detail in the 9-11 chapters and sections of this paper.This Field is it is known that the value used in the measuring method using specific sample type (for example, reference levels, cutoff value, threshold value, special Property, sensibility, caliberator and/or the concentration of control etc.) (for example, such as using the immunoassay of serum or using whole blood Point-of-care device) it techniques known in the art can be used is extrapolated to other determination forms, such as measurement standardization.For example, It is the caliberator by the way that a factor to be applied to use in measuring method that a kind of standardized mode, which can be measured, so that sample Product concentration readings are higher or lower, to obtain and the consistent slope of comparator method.The result mark that will be obtained in a kind of measuring method Other methods of standardization to another measuring method are well-known and have described in the literature (see, for example, David Wild, Immunoassay Handbook, the 4th edition, the 3.5th chapter, the 315-322 pages, content is herein incorporated by reference this Text).

3, the side of the traumatic brain injury degree in the human experimenter that may suffer from the damage to head is assisted in Method.

This disclosure relates to which a kind of assist in the traumatic brain in the doubtful human experimenter to the damage on head The side of degree of injury (such as determining whether subject suffers from mild trauma cerebral injury or moderate to severe trauma cerebral injury) Method.As used herein, " determine the traumatic brain injury degree in human experimenter " and refer to using the method (with or not companion Have other methods) determine that subject is more likely to slight TBI or moderate to severe TBI.The method includes to from tested At least two samples that person obtains are measured, and the early stage biology mark of traumatic brain injury is detected at least two samples Remember object.First sample human experimenter is derived from 24 hours of damage and the second sample after obtaining the first sample about 3 to It is derived from human experimenter within about 6 hours.Early stage biomarker is ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1).UCH-L1 is in head Portion's damage is presented in about 2 to about 24 hours after starting.The presence of UCH-L1 starts after suspicious lesion starts about 0 to about 6 hour Interior presentation.The level of UCH-L1 is determined for each of the first sample and the second sample.Determine that the horizontal of UCH-L1 is reduced Or increase.Horizontal based on UCH-L1 is to reduce, increase and be also to maintain identical from the first sample to the second sample, determines subject In traumatic brain injury degree.In the subject with traumatic brain injury, early stage, biomarker was after suspicious lesion Increase in about 0 to about 6 hour, then decreases or increases again.In some embodiments, the presence of UCH-L1 is to head injury Start to present in about 0, about 0.5 hour, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours or about 6 hours afterwards.Sample Product can be biological sample.

The method includes being measured to obtaining at least two samples from subject.First sample in suspicious lesion 24 Subject is derived from hour and the second sample is derived from subject in about 3 to about 6 hours after obtaining the first sample.The method packet The early stage biomarker that traumatic brain injury is detected at least two samples is included, the early stage biomarker is by ubiquitin carboxylic Base terminal hydrolase-l 1 (UCH-L1) composition, wherein the presence of UCH-L1 start in about 0 to about 6 hour after suspicious lesion be in It is existing；Measure the first sample and the second sample respectively in UCH-L1 it is horizontal, and determine UCH-L1 it is horizontal from the first sample to Second sample is to reduce or increase；And the level based on UCH-L1 is reduction, increases also from the first sample to the second sample It is to maintain identical, determines the traumatic brain injury degree in subject.

In some embodiments, first time point of first sample in 24 hours of suspicious lesion is derived from subject, And second time point of second sample after first time point is derived from subject and works as the horizontal from the first sample of UCH-L1 When product are reduced to the second sample, determine subject with slightly or slightly to severe trauma cerebral injury.In some embodiments In, UCH-L1 is reduced by least about 5% from increased level.For example, UCH-L1 level may from it is increased it is horizontal reduce about 5%, About 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, about 900% or about 1000%.One In a little embodiments, UCH-L1 from increased level be reduced by least about 0.1 times, at least about 0.2 times, at least about 0.3 times, at least about 0.4 times, at least about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 Again, at least about 0.9 times, at least about 1 times, at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, At least about 6 times, at least about 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times or at least 20 times.In some embodiments, UCH-L1 from it is increased it is horizontal reduce be less than about 0.1 times, less than about 0.2 times, less than about 0.3 times, less than about 0.4 times, be less than about 0.5 times, be less than about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 Times, be less than about 1 times, be less than about 1.5 times, be less than about 2 times, be less than about 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 times, it is small In about 7 times, less than 8 times, less than 9 times, less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, less than 15 Again, less than 16 times, less than 17 times, less than 18 times, less than 19 times or less than 20 times.

When the UCH-L1 level in the first sample (increases and adds deduct compared to the UCH-L1 level variation in the second sample Less) when at least about 20pg/mL at least about 6100pg/mL, subject is evaluated as with slightly or slightly to severe trauma Cerebral injury.In some embodiments, the variation (increasing or decreasing) of UCH-L1 level can be at least about 20pg/mL to about 6100pg/mL, at least about 25pg/mL are to about 6100pg/mL, at least about 30pg/mL to about 6100pg/mL, at least about 40pg/mL To about 6100pg/mL, at least about 50pg/mL to about 6100pg/mL, at least about 100pg/mL to about 6100pg/mL, at least about 129pg/mL to about 6100pg/mL, at least about 200pg/mL to about 6100pg/mL, at least about 300pg/mL to about 6100pg/ ML, at least about 400pg/mL are to about 6100pg/mL, at least about 500pg/mL to about 6100pg/mL, at least about 600pg/mL to about 6100pg/mL, at least about 700pg/mL to about 6100pg/mL, at least about 800pg/mL to about 6100pg/mL, at least about 900pg/mL to about 6100pg/mL, at least about 1000pg/mL to about 6100pg/mL, at least about 2000pg/mL to about 6100pg/ ML, at least about 3000pg/mL are to about 6100pg/mL, at least about 4000pg/mL to about 6100pg/mL, at least about 5000pg/mL To about 6100pg/mL, at least about 20pg/mL to about 4000pg/mL, at least about 25pg/mL to about 4000pg/mL, at least about 30pg/mL to about 4000pg/mL, at least about 40pg/mL to about 4000pg/mL, at least about 50pg/mL to about 4000pg/mL, extremely Few about 100pg/mL to about 4000pg/mL, at least about 129pg/mL to about 4000pg/mL, at least about 200pg/mL is to about 4000pg/mL, at least about 300pg/mL to about 4000pg/mL, at least about 400pg/mL to about 4000pg/mL, at least about 500pg/mL to about 4000pg/mL, at least about 600pg/mL to about 4000pg/mL, at least about 700pg/mL to about 4000pg/ ML, at least about 800pg/mL to about 4000pg/mL, at least about 900pg/mL to about 4000pg/mL, at least about 1000pg/mL extremely About 4000pg/mL, at least about 2000pg/mL to about 4000pg/mL, at least about 3000pg/mL to about 4000pg/mL, at least about 20pg/mL to about 2500pg/mL, at least about 25pg/mL to about 2500pg/mL, at least about 30pg/mL to about 2500pg/mL, extremely Few about 40pg/mL to about 2500pg/mL, at least about 50pg/mL to about 2500pg/mL, at least about 100pg/mL to about 2500pg/ ML, at least about 129pg/mL are to about 2500pg/mL, at least about 200pg/mL to about 2500pg/mL, at least about 300pg/mL to about 2500pg/mL, at least about 400pg/mL to about 2500pg/mL, at least about 500pg/mL to about 2500pg/mL, at least about 600pg/mL to about 2500pg/mL, at least about 700pg/mL to about 2500pg/mL, at least about 800pg/mL to about 2500pg/ ML, at least about 900pg/mL to about 2500pg/mL, at least about 1000pg/mL to about 2500pg/mL, at least about 2000pg/mL extremely About 2500pg/mL, at least about 20pg/mL are to about 1000pg/mL, at least about 25pg/mL to about 1000pg/mL, at least about 30pg/ ML to about 1000pg/mL, at least about 40pg/mL to about 1000pg/mL, at least about 50pg/mL to about 1000pg/mL, at least about 100pg/mL to about 1000pg/mL, at least about 129pg/mL to about 1000pg/mL, at least about 200pg/mL to about 1000pg/ ML, at least about 300pg/mL are to about 1000pg/mL, at least about 400pg/mL to about 1000pg/mL, at least about 500pg/mL to about 1000pg/mL, at least about 600pg/mL to about 1000pg/mL, at least about 700pg/mL to about 1000pg/mL, at least about 800pg/mL to about 1000pg/mL, at least about 900pg/mL to about 1000pg/mL, at least about 20pg/mL to about 500pg/mL, At least about 25pg/mL to about 500pg/mL, at least about 30pg/mL to about 500pg/mL, at least about 40pg/mL to about 500pg/ ML, at least about 50pg/mL are to about 500pg/mL, at least about 100pg/mL to about 500pg/mL, at least about 129pg/mL to about 500pg/mL, at least about 200pg/mL are to about 500pg/mL, at least about 300pg/mL to about 500pg/mL, at least about 400pg/mL To about 500pg/mL, at least about 20pg/mL to about 100pg/mL, at least about 25pg/mL to about 100pg/mL, at least about 30pg/ ML to about 100pg/mL, at least about 40pg/mL to about 100pg/mL, at least about 50pg/mL to about 100pg/mL, at least about 20pg/mL to about 50pg/mL, at least about 25pg/mL to about 50pg/mL, at least about 30pg/mL to about 50pg/mL or at least about 40pg/mL to about 50pg/mL.

For example, variation (increasing or decreasing) can be at least about 20pg/mL, at least about 25pg/mL, at least about 30pg/ ML, at least about 40pg/mL, at least about SOpg/mL, at least about 60pg/mL, at least about 70pg/mL, at least about 80pg/mL, at least About 90pg/mL, at least about 100pg/mL, at least about 129pg/mL, at least about 200pg/mL, at least about 300pg/mL, at least about 400pg/mL, at least about S00pg/mL, at least about 600pg/mL, at least about 700pg/mL, at least about 800pg/mL, at least about 900pg/mL, at least about 1000pg/mL, at least about 1500pg/mL, at least about 2000pg/mL, at least about 2528pg/mL, at least About 2500pg/mL, at least about 3000pg/mL, at least about 3500pg/mL, at least about 4000pg/mL, at least about 4500pg/mL, At least about 5000pg/mL, at least about 5500pg/mL, at least about 6000pg/mL or at least about 6100pg/mL.

In some embodiments, the first sample obtains in 24 hours after suspicious lesion.For example, the first sample can be After suspicious lesion in about 0 hour, in about 1 hour, in about 2 hours, in about 3 hours, in about 4 hours, in about 5 hours, about 6 hours It is interior, in about 7 hours, in about 8 hours, in about 9 hours, in about 10 hours, in about 11 hours, in about 12 hours, in about 13 hours, In about 14 hours, in about 15 hours, in about 16 hours, in about 17 hours, in about 18 hours, in about 19 hours, in about 20 hours, It is obtained in about 21 hours, in about 22 hours, in about 23 hours, in about 24 hours or more than about 24 hours.In some embodiments In, the first sample can after suspicious lesion in about 0 to about 1 hour, in about 0 to about 2 hour, in about 0 to about 3 hour, about 0 to In about 4 hours, in about 0 to about 5 hour, in about 0 to about 6 hour, in about 0 to about 7 hour, in about 0 to about 8 hour, about 0 to about In 9 hours, in about 0 to about 10 hour, in about 0 to about 11 hour, in about 0 to about 12 hour, in about 0 to about 18 hour, about 6 to It is obtained in about 12 hours, in about 12 to about 18 hours, in about 18 to about 24 hours or greater than 24 hours.

In some embodiments, the second sample is about 1 hour to about 10 hours, such as the first sample after the first sample obtains Product obtain in about 3 hours to about 6 hours after obtaining.In some embodiments, the second sample is about 1 small after the acquisition of the first sample When, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours or about 10 hours Interior acquisition.

In some embodiments, the first sample obtains and works as the water of UCH-L1 after suspicious lesion in 0 to about 6 hour It is flat when increasing or decreasing at least about 20pg/mL at least about 6100pg/mL from the first sample to the second sample, determine that subject suffers from There is moderate to severe trauma cerebral injury.

In some embodiments, the first sample can obtain and the increase of UCH-L1 level in about 0 to about 6 hour Or it is reduced at least about 2528pg/mL.In some embodiments, the first sample can be obtained in about 0 to about 6 hour and UCH-L1 is horizontal to be increased or decreased as at least about 129pg/mL.In some embodiments, the first sample can be about 0 to about It is obtained in 10 hours and UCH-L1 is horizontal increases or decreases as at least about 25pg/mL.In some embodiments, the first sample Product can obtain in about 0 to about 11 hour and UCH-L1 is horizontal increases or decreases as at least about 25pg/mL.In some realities It applies in mode, the first sample can obtain in about 0 to about 12 hour and UCH-L1 is horizontal increases or decreases as at least about 129pg/mL。

In some embodiments, when interior obtained from subject's when after to head injury or suspicious lesion about 0 to about 6 is small The amount of the UCH-L1- antibody complex detected in one or more samples increases to rise, and (for example, about 100pg/mL is to about When 1000pg/mL), it is believed that subject suffers from moderate to severe trauma cerebral injury.In some embodiments, UCH-L1 Level can be about 0 to about 6 hour, about 0.5 hour to about 6 hours, about 1 hour to about 6 hours, about 1.5 after to head injury Hour to about 6 hours, about 2 hours to about 6 hours, about 2.5 hours to about 6 hours, about 3 hours to about 6 hours, about 4 hours extremely About 6 hours, about 5 hours to about 6 hours, about 0 to about 5 hour, about 0.5 hour to about 5 hours, about 1 hour to about 5 hours, about 1.5 hours to about 5 hours, about 2 hours to about 5 hours, about 2.5 hours to about 5 hours, about 3 hours to about 5 hours, about 4 hours To about 5 hours, about 0 to about 4 hour, about 0.5 hour to about 4 hours, about 1 hour to about 4 hours, it is about 1.5 hours to about 4 small When, about 2 hours to about 4 hours, about 2.5 hours to about 4 hours, about 3 hours to about 4 hours, about 0 to about 3 hour, it is about 0.5 small Up to about 3 hours, about 1 hour to about 3 hours, about 1.5 hours to about 3 hours, about 2 hours to about 3 hours, about 2.5 hours extremely About 3 hours, about 0 to about 2.5 hour, about 0.5 hour to about 2.5 hours, about 1 hour to about 2.5 hours, about 1.5 hours to about 2.5 hour, about 2 hours to about 2.5 hours, about 0 to about 2 hour, about 0.5 hour to about 2 hours, about 1 hour to about 2 hours or Increase in about 1.5 hours to about 2 hours.In some embodiments, UCH-L1 level can about 0 after to head injury, about 0.5 hour, about 1 hour, about 1.5 hours, about 2 hours, about 2.5 hours, about 3 hours, about 3.5 hours, about 4 hours, it is about 4.5 small When, about 5 hours, increase in about 5.5 hours or about 6 hours.

In some embodiments, after to head injury or suspicious lesion, UCH-L1 level can increase to rise. The rise of UCH-L1 can be at least about 20pg/mL between at least about 25000pg/mL.For example, the rise of UCH-L1 can With at least about 20pg/mL at least about 25000pg/mL, at least about 50pg/mL at least about 25000pg/mL, at least about 100pg/mL at least about 25000pg/mL, at least about 200pg/mL at least about 25000pg/mL, at least about 300pg/mL is extremely At least about 25000pg/mL, at least about 400pg/mL are at least about 25000pg/mL, at least about 500pg/mL at least about 25000pg/mL, at least about 1000pg/mL are at least about 25000pg/mL, at least about 2000pg/mL at least about 25000pg/ ML, at least about 3000pg/mL at least about 25000pg/mL, at least about 4000pg/mL at least about 25000pg/mL, at least about 5000pg/mL at least about 25000pg/mL, at least about 20pg/mL at least about 15000pg/mL, at least about 50pg/mL is to extremely Few about 15000pg/mL, at least about 100pg/mL at least about 15000pg/mL, at least about 200pg/mL is at least about 15000pg/mL, at least about 300pg/mL at least about 15000pg/mL, at least about 400pg/mL at least about 15000pg/mL, At least about 500pg/mL at least about 15000pg/mL, at least about 1000pg/mL at least about 15000pg/mL, at least about 2000pg/mL at least about 15000pg/mL, at least about 3000pg/mL is at least about 15000pg/mL, at least about 4000pg/mL To at least about 15000pg/mL, at least about 5000pg/mL at least about 15000pg/mL, at least about 10000pg/mL at least about 15000pg/mL, at least about 20pg/mL at least about 10000pg/mL, at least about 50pg/mL at least about 10000pg/mL, extremely Lack about 100pg/mL at least about 10000pg/mL, at least about 200pg/mL at least about 10000pg/mL, at least about 300pg/ ML at least about 10000pg/mL, at least about 400pg/mL at least about 10000pg/mL, at least about 500pg/mL is at least about 10000pg/mL, at least about 1000pg/mL are at least about 10000pg/mL, at least about 2000pg/mL at least about 10000pg/ ML, at least about 3000pg/mL at least about 10000pg/mL, at least about 4000pg/mL at least about 10000pg/mL, at least about 5000pg/mL at least about 10000pg/mL, at least about 20pg/mL at least about 5000pg/mL, at least about 50pg/mL is to extremely Few about 5000pg/mL, at least about 100pg/mL at least about 5000pg/mL, at least about 200pg/mL at least about 5000pg/ ML, at least about 300pg/mL at least about 5000pg/mL, at least about 400pg/mL at least about 5000pg/mL, at least about 500pg/mL at least about 5000pg/mL, at least about 1000pg/mL at least about 5000pg/mL, at least about 2000pg/mL is extremely At least about 5000pg/mL, at least about 3000pg/mL are at least about 5000pg/mL, at least about 4000pg/mL at least about 5000pg/mL, at least about 20pg/mL at least about 1000pg/mL, at least about 50pg/mL at least about 1000pg/mL, at least About 100pg/mL at least about 1000pg/mL, at least about 200pg/mL at least about 1000pg/mL, at least about 300pg/mL is extremely At least about 1000pg/mL, at least about 400pg/mL are at least about 1000pg/mL or at least about 500pg/mL at least about Between 1000pg/mL.In some embodiments, the rise of UCH-L1 can be at least about 100pg/mL, at least about 200pg/mL, at least about 300pg/mL, at least about 400pg/mL, at least about 500pg/mL, at least about 600pg/mL, at least about 700pg/mL, at least about 800pg/mL, at least about 900pg/mL, at least about 1000pg/mL, at least about 21000pg/mL, at least About 3000pg/mL, at least about 4000pg/mL, at least about 5000pg/mL, at least about 10000pg/mL or at least about 15000pg/ mL。

In some embodiments, the method further includes being assessed as suffering from the treatment of traumatic brain injury therapy Moderate to severe trauma cerebral injury subject, as described below.In some embodiments, the method further includes prisons It surveys and is assessed as the subject with mild trauma cerebral injury, as described below.

The property of the measuring method used in methods described herein is not critical, and is tested and be can be known in the art What measuring method, such as immunoassay, the Western Immuno precipitation method, immunoelectrophoresis, chemical analysis, SDS-PAGE and egg White matter blot analysis or Western Immuno decoration method, electrophoretic analysis, protein determination, competitive binding assay, function Energy property protein determination or chromatography or spectroscopic methodology, such as high performance liquid chromatography (HPLC) or liquid chromatography-mass spectrography (LC/ MS).In addition, measuring method can be used in the form of clinical chemistry, it such as will be by known to persons of ordinary skill in the art.Such survey Determine method to be described in further detail in the 9-11 chapters and sections of this paper.

4, the method for evaluating whether to carry out CT scan to the human experimenter for suffering from the damage to head

In addition to other methods, this disclosure relates to which one kind assists in or evaluates whether to suffering from or may suffer from doubtful The damage to head human experimenter carry out computerized tomography (CT) scanning method.As used herein, " determine whether pair Human experimenter's progress CT scan ", which refers to using the method (with or without other methods), determines that subject is more likely to With with positive head CT scan.Specifically, the method can comprise the following steps that (a) is obtained at least to from subject Two samples are measured, and the first sample is derived from subject in 24 hours of suspicious lesion, and the second sample is obtaining the It is derived from subject within about 3 to about 6 hours after a sample；(b) biology early stage detecting traumatic brain injury at least two samples Marker, the early stage biomarker are made of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1), the wherein presence of UCH-L1 Start to present in about 0 to about 6 hour after suspicious lesion；(c) measure the first sample and the second sample respectively in UCH-L1 water It is flat, and determine that the level of UCH-L1 is reduction from the first sample to the second sample or increases；And (d) based on UCH-L1's Level from the first sample to the second sample be reduce, increase also be to maintain it is identical, it is determined whether to subject carry out CT scan.In In some embodiments, when the UCH-L1 level in the first sample or the second sample be higher than UCH-L1 reference levels when, to by Examination person carries out CT scan.In some embodiments, when the UCH-L1 level in the first sample or the second sample is lower than UCH-L1 Reference levels when, not to subject carry out CT scan.In some embodiments, when horizontal from the UCH-L1 in the first sample When increasing or decreasing, CT scan is carried out to subject with statistically significant to the UCH-L1 level in the second sample.In In some embodiments, when no statistically from the UCH-L1 level in the first sample to the UCH-L1 level in the second sample When significantly increasing or decreasing, CT scan is not carried out to subject.In some embodiments, horizontal from first as UCH-L1 When sample decreases or increases at least absolute magnitude to the second sample, CT scan is carried out to subject.In some embodiments, when When the level of UCH-L1 does not increase or decrease at least absolute magnitude from the first sample to the second sample, CT is not carried out to subject and swept It retouches.In some embodiments, when the UCH-L1 level in the first sample is higher than the reference levels of UCH-L1 or works as from the first sample UCH-L1 level in product have to the UCH-L1 level in the second sample it is statistically significant when increasing or decreasing, to tested Person carries out CT scan.In some embodiments, when the UCH-L1 level in the first sample lower than UCH-L1 reference levels or It adds deduct when from the UCH-L1 level in the first sample to the not statistically significant increasing of the UCH-L1 level in the second sample When few, CT scan is not carried out to subject.Sample can be biological sample.

In some embodiments, subject received CT scan before or after being measured.In some embodiment party In formula, it is based on CT scan, subject is doubtful to suffer from traumatic brain injury.In some embodiments, by reference levels and the positive Head CT scan is associated.

In general, reference levels may be used as the benchmark of the result obtained when the UCH-L1 of assessment measurement test sample. In general, when carrying out this compare, reference levels obtain in the following manner: transporting with enough numbers and under proper condition Row particular assay method so that can by the moment or terminal of analyte existence, amount or concentration and TBI or with specific mark Will is associated or is associated with.In general, obtaining reference levels with the measurement to reference subject (or subject group).It is surveyed The UCH-L1 of amount may include UCH-L1, its segment, its catabolite and/or its enzymatic lysis product.

In some embodiments, reference levels be by between at least about 80% to about 100% sensibility and What the measuring method of the specificity between at least about 30% to about 100% determined.In some embodiments, sensibility is at least about 80.0%, at least about 85.0%, at least about 87.5%, at least about 90.0%, at least about 95.0%, at least about 99.0%, at least About 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, extremely Few about 99.7%, at least about 99.8%, at least about 99.9% or at least about 100.0%.In some embodiments, specificity is At least about 30.0%, at least about 31.0%, at least about 32.0%, at least about 33.0%, at least about 34.0%, at least about 35.0%, at least about 36.0%, at least about 37.0%, at least about 38.0%, at least about 39.0%, at least about 40.0%, at least About 45.0%, at least about 50.0%, at least about 55.0%, at least about 60.0%, at least about 65.0%, at least about 70.0%, extremely Few about 75.0%, at least about 80.0%, at least about 85.0%, at least about 90.0%, at least about 91.0%, at least about 92.0%, At least about 93.0%, at least about 94.0%, at least about 95.0%, at least about 96.0%, at least about 97.0%, at least about 98.0%, at least about 99.0%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least About 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9% or at least about 100.0%. For example, sensibility is at least about 99% and specificity is at least about 75%, sensibility is at least about 99% and specificity is At least about 99% or sensibility is about 100% and specificity is about 100%

In some embodiments, reference levels can be at least about 50pg/mL between about 1000pg/mL.Some In embodiment, reference levels can at least about 50pg/mL between about 1000pg/mL, at least about 50pg/mL to about Between 900pg/mL, at least about 50pg/mL between about 800pg/mL, at least about 50pg/mL between about 750pg/mL, At least about 50pg/mL between about 700pg/mL, at least about 50pg/mL between about 600pg/mL, at least about 50pg/ ML between about 500pg/mL, at least about 50pg/mL between about 400pg/mL, at least about 50pg/mL to about 300pg/ Between mL, at least about 50pg/mL between about 200pg/mL, at least about 100pg/mL between about 1000pg/mL, extremely Few about 100pg/mL between about 900pg/mL, at least about 100pg/mL between about 800pg/mL, at least about 100pg/ ML between about 750pg/mL, at least about 100pg/mL between about 700pg/mL, at least about 100pg/mL to about Between 600pg/mL, at least about 100pg/mL between about 500pg/mL, at least about 100pg/mL to about 400pg/mL it Between, at least about 100pg/mL between about 300pg/mL, at least about 100pg/mL between about 200pg/mL, at least about 150pg/mL between about 1000pg/mL, at least about 150pg/mL between about 900pg/mL, at least about 150pg/mL extremely Between about 800pg/mL, at least about 150pg/mL between about 750pg/mL, at least about 150pg/mL to about 700pg/mL Between, at least about 150pg/mL between about 600pg/mL, at least about 150pg/mL between about 500pg/mL, at least About 150pg/mL between about 400pg/mL, at least about 150pg/mL between about 300pg/mL, at least about 150pg/mL To between about 200pg/mL, at least about 200pg/mL between about 1000pg/mL, at least about 200pg/mL to about 900pg/ Between mL, at least about 200pg/mL between about 800pg/mL, at least about 200pg/mL between about 750pg/mL, extremely Few about 200pg/mL between about 700pg/mL, at least about 200pg/mL between about 600pg/mL, at least about 200pg/ ML between about 500pg/mL, at least about 200pg/mL between about 400pg/mL, at least about 200pg/mL to about Between 300pg/mL, at least about 300pg/mL between about 1000pg/mL, at least about 300pg/mL to about 900pg/mL it Between, at least about 300pg/mL between about 800pg/mL, at least about 300pg/mL between about 750pg/mL, at least about 300pg/mL between about 700pg/mL, at least about 300pg/mL between about 600pg/mL, at least about 300pg/mL extremely Between about 500pg/mL, at least about 300pg/mL between about 400pg/mL, at least about 400pg/mL to about 1000pg/mL Between, at least about 400pg/mL between about 900pg/mL, at least about 400pg/mL between about 800pg/mL, at least About 400pg/mL between about 750pg/mL, at least about 400pg/mL between about 700pg/mL, at least about 400pg/mL To between about 600pg/mL, at least about 400pg/mL between about 500pg/mL, at least about 500pg/mL to about 1000pg/ Between mL, at least about 500pg/mL between about 900pg/mL, at least about 500pg/mL between about 800pg/mL, extremely Few about 500pg/mL between about 750pg/mL, at least about 500pg/mL between about 700pg/mL or at least about 500pg/ ML is between about 600pg/mL.For example, reference levels can be at least about 50pg/mL, 55pg/mL, 60pg/mL, 65pg/mL, 70pg/mL、75pg/mL、80pg/mL、85pg/mL、90pg/mL、95pg/mL、96pg/mL、97pg/mL、98pg/mL、98pg/ mL、100pg/mL、150pg/mL、200pg/mL、250pg/mL、300pg/mL、350pg/mL、370pg/mL、400pg/mL、 450pg/mL, 500pg/mL, 509pg/mL 550pg/mL, 600pg/mL, 700pg/mL, 800pg/mL, 900pg/mL or 1000pg/mL。

In some embodiments, the first sample obtains in 24 hours after suspicious lesion.For example, the first sample can be After suspicious lesion in about 0 hour, in about 30 minutes, in about 1 hour, in about 2 hours, in about 3 hours, in about 4 hours, it is about 5 small When it is interior, in about 6 hours, in about 7 hours, in about 8 hours, in about 9 hours, in about 10 hours, in about 11 hours, about 12 hours It is interior, in about 13 hours, in about 14 hours, in about 15 hours, in about 16 hours, in about 17 hours, in about 18 hours, about 19 hours It is interior, obtain in about 20 hours, in about 21 hours, in about 22 hours, in about 23 hours, in about 24 hours or more than about 24 hours. In some embodiments, the first sample after suspicious lesion in about 0 to about 6 hour, in about 6 to about 12 hours, about 12 to about In 18 hours, about 18 to about 24 hours or be greater than 24 hours obtain.

In some embodiments, the first sample obtains in 24 hours after suspicious lesion, and reference levels are to pass through What the measuring method with certain sensitivity as described above and specificity determined.In some embodiments, sample is in doubtful damage After wound in about 0 to about 6 hour, in about 0 to about 7 hour, in about 0 to about 8 hour, in about 0 to about 9 hour, about 0 to about 10 hour It is interior, in about 0 to about 11 hour, in about 0 to about 12 hour, in about 6 to 12 hours, in about 12 to about 18 hours, about 18 to about 24 It is obtained in hour or greater than 24 hours and reference levels is by with certain sensitivity and specificity such as about 100% Sensibility and at least about 75% specificity, at least about 99% sensibility and at least about 75% specificity or about 100% Sensibility and about 100% specificity measuring method determine.

In some embodiments, the first sample can obtain and join between after suspicious lesion about 0 hour to 6 hours Level is examined to be at least about 370pg/mL and be the survey by having the specificity of about 100% sensibility and at least about 37.5% Determine what method determined.In some embodiments, the first sample obtains and joins between after suspicious lesion about 0 hour to 6 hours The level examined is about 509pg/mL and is true by the measuring method with the specificity of about 100% sensibility and at least about 75% Fixed.In some embodiments, the first sample obtains and reference levels between after suspicious lesion about 6 hours to 12 hours It is about 96pg/mL and is determined by the measuring method with the specificity of at least about 96% sensibility and at least about 30% 's.In some embodiments, the first sample obtains between after suspicious lesion about 6 hours to 12 hours and reference levels are About 86pg/mL and be by at least about 86% sensibility and at least about 35% specificity measuring method determination.

In some embodiments, absolute magnitude can be by having the sensibility between at least about 80% to about 100% What the measuring method of the specificity between at least about 30% to about 100% determined.In some embodiments, sensibility is at least About 80.0%, at least about 85.0%, at least about 87.5%, at least about 90.0%, at least about 95.0%, at least about 99.0%, extremely Few about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, At least about 99.7%, at least about 99.8%, at least about 99.9% or at least about 100.0%.In some embodiments, specific For at least about 30.0%, at least about 31.0%, at least about 32.0%, at least about 33.0%, at least about 34.0%, at least about 35.0%, at least about 36.0%, at least about 37.0%, at least about 38.0%, at least about 39.0%, at least about 40.0%, at least About 45.0%, at least about 50.0%, at least about 55.0%, at least about 60.0%, at least about 65.0%, at least about 70.0%, extremely Few about 75.0%, at least about 80.0%, at least about 85.0%, at least about 90.0%, at least about 91.0%, at least about 92.0%, At least about 93.0%, at least about 94.0%, at least about 95.0%, at least about 96.0%, at least about 97.0%, at least about 98.0%, at least about 99.0%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least About 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9% or at least about 100.0%. For example, sensibility can be at least about 99% and specificity can be at least about 75%, sensibility can be at least about 99% And specificity can be at least about 99% or sensibility can be about 100% and specificity can be about 100%

In some embodiments, sample after suspicious lesion in 0 to 10 hour obtain and absolute magnitude such as about 25pg/ ML is determined by the measuring method with the specificity of at least about 85% sensibility and at least about 41%.In some embodiment party In formula, sample obtains in 0 to 10 hour after suspicious lesion and absolute magnitude is about 25pg/mL.In some embodiments, sample Product obtain in 0 to 10 hour after suspicious lesion and absolute magnitude, such as about 23pg/mL, are by having at least about 90% Sensibility and at least about 35% specificity measuring method determine.In some embodiments, sample after suspicious lesion 0 to It is obtained in 10 hours and absolute magnitude is about 23pg/mL.

In some embodiments, when the UCH-L1 level from the first sample that first time point obtains is to second The UCH-L1 level in the second sample that time point obtains when increasing or decreasing, carries out subject with statistically significant CT scan.In some embodiments, when from the UCH-L1 level in the first sample to the second sample obtained at the second time point UCH-L1 level in product it is not statistically significant when increasing or decreasing, CT scan is not carried out to subject.In some realities It applies in mode, from the UCH-L1 level in the first sample that first time point obtains to the second sample obtained at the second time point Statistically significant the increasing or decreasing of UCH-L1 level in product is to be more than about 0.1 times, be more than about 0.2 times, more than about 0.3 Times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, more than about 0.6 times, more than about 0.7 times, more than about 0.73 times, More than about 0.8 times, more than about 0.9 times, more than about 1 times, be more than 1.5 times, be more than 1.81 times, more than about 2 times, be more than about 3 times, surpass Cross about 4 times, more than about 5 times, more than about 6 times, more than about 7 times, more than 8 times, more than 9 times, more than 10 times, more than 11 times, be more than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 times, more than 18 times, more than 19 times or more than 20 Times.In some embodiments, from the UCH-L1 level in the first sample that first time point obtains at the second time point Horizontal statistically significant of the UCH-L1 in the second sample obtained reduce by more than about 0.1 times, more than about 0.2 times, be more than about 0.3 times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, more than about 0.6 times, more than about 0.7 times, be more than about 0.73 Again, more than about 0.8 times, more than about 0.9 times, more than about 1 times, more than 1.5 times, more than 1.81 times, more than about 2 times, more than about 3 Times, more than about 4 times, more than about 5 times, more than about 6 times, more than about 7 times, more than about 8 times, more than 9 times, more than 10 times, more than 11 Times, more than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 times, more than 18 times, more than 19 times or Show that subject should carry out CT scan more than 20 times.In some embodiments, from the first sample obtained in first time point In UCH-L1 level to the UCH-L1 level in the second sample that the second time point obtained statistically significant increase it is super Cross about 0.1 times, more than about 0.2 times, more than about 0.3 times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, be more than about 0.6 times, more than about 0.7 times, more than about 0.73 times, more than about 0.8 times, more than about 0.9 times, more than about 1 times, more than about 1.5 times, More than 1.81 times, more than about 2 times, more than about 3 times, more than about 4 times, more than about 5 times, more than about 6 times, more than about 7 times, more than 8 Times, be more than 9 times, more than 10 times, more than 11 times, more than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, surpass It crosses 17 times, show that subject should carry out CT scan more than 18 times, more than 19 times or more than 20 times.

In some embodiments, from the UCH-L1 level in the first sample that first time point obtains at second Between put statistically significant the increasing or decreasing of the UCH-L1 level in the second sample of acquirement and be less than about 0.1 times, be less than About 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, be less than about 1 times, less than 1.5 times, less than 1.81 times, it is small In about 2 times, be less than about 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 times, be less than about 7 times, less than 8 times, less than 9 times, it is small In 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 times, less than 18 Again, less than 19 times or less than 20 times.In some embodiments, the UCH-L1 from the first sample that first time point obtains Level to the UCH-L1 level in the second sample that the second time point obtained statistically significant reduction be less than about 0.1 times, Less than about 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least About 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, be less than about 1 times, less than 1.5 times, less than 1.81 times, Less than about 2 times, be less than about 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 times, be less than about 7 times, be less than about 8 times, less than 9 Times, less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 times, it is small Show that subject should carry out CT scan in 18 times, less than 19 times or less than 20 times.In some embodiments, from the first time System of the UCH-L1 level in the first sample that point obtains to the UCH-L1 level in the second sample that the second time point obtained Meter learn on dramatically increase less than about 0.1 times, be less than about 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, extremely Lack about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, be less than About 1 times, be less than about 1.5 times, less than 1.81 times, be less than about 2 times, be less than about 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 Times, be less than about 7 times, less than 8 times, less than 9 times, less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, it is small Show that subject should carry out CT scan in 15 times, less than 16 times, less than 17 times, less than 18 times, less than 19 times or less than 20 times.In In some embodiments, from the UCH-L1 level in the first sample that first time point obtains to obtaining at the second time point Not statistically significant the increasing or decreasing of UCH-L1 level in second sample shows that subject does not need to carry out CT scan.

In some embodiments, first time point is about 0 to about 12 hour after suspicious lesion and statistically aobvious Increasing or decreasing for writing is to be less than about 2 times from first time point to the second time point.In some embodiments, first time point It is about 0 to about 12 hour after suspicious lesion and when statistically significant increasing or decreasing is from first time point to second Between point be less than about 1.81 times.In some embodiments, first time point is about 0 to about 12 hour after suspicious lesion and unites It is from the second time point to first time point more than about 0.50 times that meter, which learns upper significant increase,.In some embodiments, first When time point is about 0 to about 12 hour after suspicious lesion and statistically significant increase is from the second time point to first Between point be more than about 0.55 times.

In some embodiments, the method further includes with traumatic brain injury therapy treat subject and/or Subject is monitored, as described below.

The property of the measuring method used in methods described herein is not critical, and test can be it is as known in the art Any measuring method, such as immunoassay, the Western Immuno precipitation method, immunoelectrophoresis, chemical analysis, SDS-PAGE and Western blot analysis or Western Immuno decoration method, electrophoretic analysis, protein determination, competitive binding assay, function Energy property protein determination or chromatography or spectroscopic methodology, such as high performance liquid chromatography (HPLC) or liquid chromatography-mass spectrography (LC/ MS).In addition, measuring method can be used in the form of clinical chemistry, it such as will be by known to persons of ordinary skill in the art.Such survey Determine method to be described in further detail in the 9-11 chapters and sections of this paper.

5, predict the doubtful human experimenter with head injury whether by the side with positive or negative head CT scan Method

In addition to other methods, this disclosure relates to a kind of prediction or the doubtful mankind with the damage to head of aid forecasting by Whether examination person is by the method with positive or negative head CT scan.As used herein, it " predicts whether to carry out human experimenter CT scan ", which refers to using the method (with or without other methods), predicts that subject is more likely to with positive head Portion's CT scan.The described method includes: (a) is measured the first sample and the second optional sample, with measurement or test sample The level of middle ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1), wherein the first sample after suspicious lesion in 24 hours first when Between point be derived from subject and second time point of second sample after first time point, about 3 such as after first time point Hour was derived from subject to about 6 hours；(b) the early stage biology mark of traumatic brain injury is detected in the first and/or second sample Remember that object, the early stage biomarker are made of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1)；And (c) it is based on UCH-L1 water It is flat to determine whether subject have positive or negative head CT scan.(i) as the UCH- in the first sample and/or the second sample When L1 level is higher than the reference levels of UCH-L1, subject is determined as to there may be positive head CT scan, and when first When UCH-L1 level in sample and/or the second sample is lower than the reference levels of UCH-L1, subject is determined as to have Negative head CT scan；(ii) when from the UCH-L1 level in the first sample to the UCH-L1 level in the second sample have statistics When significantly increasing or decreasing on, subject is determined as to there may be positive head CT scan, and is worked as from the first sample In UCH-L1 level to the UCH-L1 level in the second sample it is not statistically significant when increasing or decreasing, by subject It is determined as there may be negative head CT scan；(iii) when the level of UCH-L1 reduces or increases from the first sample to the second sample When adding to few absolute magnitude, subject is determined as to there may be positive head CT scan, and works as the horizontal from first of UCH-L1 When sample does not decrease or increase at least absolute magnitude to the second sample, subject is determined as to there may be negative Cranial Computed Tomography to sweep It retouches；Or (iv) when the level of the UCH-L1 in the first sample is higher than the reference levels of UCH-L1, or when from the first sample UCH-L1 level to the UCH-L1 level in the second sample have it is statistical dramatically increase or reduce when, subject is true Positive head CT scan may be had by being set to, and when the level of the UCH-L1 in the first sample is lower than the reference levels of UCH-L1 When, or ought not there is no statistical significant increasing from the UCH-L1 level in the first sample to the UCH-L1 level in the second sample When adding deduct few, subject is determined as to there may be negative head CT scan.Sample can be biological sample.

In some embodiments, the first sample and/or the second sample after suspicious lesion about 2 hours to about 12 hours it Between be derived from subject.For example, the first sample and/or the second sample are about 2 hours to about 12 hours after to head injury, about 2 small Up to about 10 hours, about 2 hours to about 8 hours, about 2 hours to about 6 hours, about 2 hours to about 4 hours, about 4 hours to about 12 Hour, about 4 hours to about 10 hours, about 4 hours to about 8 hours, about 4 hours to about 6 hours, about 6 hours to about 12 hours, about It is obtained between about 10 hours or about 6 hours to about 8 hours within 6 hours.In some embodiments, the first sample and/or second Sample about 2 hours after injury, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, It obtains within about 10 hours, about 11 hours or about 12 hours.

In some embodiments, when the UCH-L1 level in the first sample and/or the second sample greater than or equal to about When 100pg/mL or 300pg/mL, subject is evaluated as to may have positive head CT scan, and when the first sample and/or When UCH-L1 level in second sample is less than about 100pg/mL or 300pg/mL, subject is evaluated as not with any wound Property cerebral injury.In some embodiments, when the UCH-L1 level in the first sample and/or the second sample greater than or equal to about 50pg/mL, about 100pg/mL, about 150pg/mL, about 200pg/mL, about 250pg/mL, about 300pg/mL, about 350pg/mL, about When 400pg/mL, about 450pg/mL or about 500pg/mL, subject is determined as to there may be positive head CT scan.Some In embodiment, when the UCH-L1 level in the first sample and/or the second sample is less than about 50pg/mL, about 100pg/mL, about 150pg/mL, about 200pg/mL, about 250pg/mL, about 300pg/mL, about 350pg/mL, about 400pg/mL, about 450pg/mL or When about 500pg/mL, subject is determined as to there may be negative head CT scan.

In some embodiments, reference levels can be by having the sensitivity between at least about 80% to about 100% Property and at least about 30% to about 100% between specificity measuring method determine.In some embodiments, sensibility is extremely Few about 80.0%, at least about 85.0%, at least about 87.5%, at least about 90.0%, at least about 95.0%, at least about 99.0%, At least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9% or at least about 100.0%.In some embodiments In, specificity at least about 30.0%, at least about 31.0%, at least about 32.0%, at least about 33.0%, at least about 34.0%, At least about 35.0%, at least about 36.0%, at least about 37.0%, at least about 38.0%, at least about 39.0%, at least about 40.0%, at least about 45.0%, at least about 50.0%, at least about 55.0%, at least about 60.0%, at least about 65.0%, at least About 70.0%, at least about 75.0%, at least about 80.0%, at least about 85.0%, at least about 90.0%, at least about 91.0%, extremely Few about 92.0%, at least about 93.0%, at least about 94.0%, at least about 95.0%, at least about 96.0%, at least about 97.0%, At least about 98.0%, at least about 99.0%, at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5%, at least about 99.6%, at least about 99.7%, at least about 99.8%, at least about 99.9% or at least About 100.0%.For example, sensibility is at least about 99% and specificity is at least about 75%, sensibility is at least about 99% simultaneously And specificity is at least about 99% or sensibility is about 100% and specificity is about 100%

In some embodiments, reference levels can be at least about 50pg/mL between about 1000pg/mL.Some In embodiment, reference levels can at least about 50pg/mL between about 1000pg/mL, at least about 50pg/mL to about Between 900pg/mL, at least about 50pg/mL between about 800pg/mL, at least about 50pg/mL between about 750pg/mL, At least about 50pg/mL between about 700pg/mL, at least about 50pg/mL between about 600pg/mL, at least about 50pg/ ML between about 500pg/mL, at least about 50pg/mL between about 400pg/mL, at least about 50pg/mL to about 300pg/ Between mL, at least about 50pg/mL between about 200pg/mL, at least about 100pg/mL between about 1000pg/mL, extremely Few about 100pg/mL between about 900pg/mL, at least about 100pg/mL between about 800pg/mL, at least about 100pg/ ML between about 750pg/mL, at least about 100pg/mL between about 700pg/mL, at least about 100pg/mL to about Between 600pg/mL, at least about 100pg/mL between about 500pg/mL, at least about 100pg/mL to about 400pg/mL it Between, at least about 100pg/mL between about 300pg/mL, at least about 100pg/mL between about 200pg/mL, at least about 150pg/mL between about 1000pg/mL, at least about 150pg/mL between about 900pg/mL, at least about 150pg/mL extremely Between about 800pg/mL, at least about 150pg/mL between about 750pg/mL, at least about 150pg/mL to about 700pg/mL Between, at least about 150pg/mL between about 600pg/mL, at least about 150pg/mL between about 500pg/mL, at least About 150pg/mL between about 400pg/mL, at least about 150pg/mL between about 300pg/mL, at least about 150pg/mL To between about 200pg/mL, at least about 200pg/mL between about 1000pg/mL, at least about 200pg/mL to about 900pg/ Between mL, at least about 200pg/mL between about 800pg/mL, at least about 200pg/mL between about 750pg/mL, extremely Few about 200pg/mL between about 700pg/mL, at least about 200pg/mL between about 600pg/mL, at least about 200pg/ ML between about 500pg/mL, at least about 200pg/mL between about 400pg/mL, at least about 200pg/mL to about Between 300pg/mL, at least about 300pg/mL between about 1000pg/mL, at least about 300pg/mL to about 900pg/mL it Between, at least about 300pg/mL between about 800pg/mL, at least about 300pg/mL between about 750pg/mL, at least about 300pg/mL between about 700pg/mL, at least about 300pg/mL between about 600pg/mL, at least about 300pg/mL extremely Between about 500pg/mL, at least about 300pg/mL between about 400pg/mL, at least about 400pg/mL to about 1000pg/mL Between, at least about 400pg/mL between about 900pg/mL, at least about 400pg/mL between about 800pg/mL, at least About 400pg/mL between about 750pg/mL, at least about 400pg/mL between about 700pg/mL, at least about 400pg/mL To between about 600pg/mL, at least about 400pg/mL between about 500pg/mL, at least about 500pg/mL to about 1000pg/ Between mL, at least about 500pg/mL between about 900pg/mL, at least about 500pg/mL between about 800pg/mL, extremely Few about 500pg/mL between about 750pg/mL, at least about 500pg/mL between about 700pg/mL or at least about 500pg/ ML is between about 600pg/mL.For example, reference levels can at least about 50pg/mL, 55pg/mL, 60pg/mL, 65pg/mL, 70pg/mL、75pg/mL、80pg/mL、85pg/mL、90pg/mL、95pg/mL、96pg/mL、97pg/mL、98pg/mL、98pg/ mL、100pg/mL、150pg/mL、200pg/mL、250pg/mL、300pg/mL、350pg/mL、370pg/mL、400pg/mL、 450pg/mL, 500pg/mL, 509pg/mL 550pg/mL, 600pg/mL, 700pg/mL, 800pg/mL, 900pg/mL or 1000pg/mL。

In some embodiments, first time point acquisition and second of first sample in 24 hours of suspicious lesion Second time point or optional third time point or fourth time point of the sample after first time point obtain, tested with determination Whether person will have positive or negative head CT scan.In some embodiments, first time point is to head injury or to doubt Like about 0 to about 24 hour after damage.For example, first time point can be about 0 to about 24 hour after suspicious lesion, about 0 To about 20 hours, about 0 to about 18 hour, about 0 to about 16 hour, about 0 to about 14 hour, about 0 to about 12 hour, about 0 to about 10 Hour, about 0 to about 8 hour, about 0 to about 6 hour, about 0 to about 4 hour, about 0 to about 2 hour, about 0 to about 1 hour, about 0 are to about 1.5 hours, about 0.5 hour to about 24 hours, about 0.5 hour to about 20 hours, about 0.5 hour to about 18 hours, about 0.5 hour To about 16 hours, about 0.5 hour to about 14 hours, about 0.5 hour to about 12 hours, about 0.5 hour to about 10 hours, about 0.5 Hour was to about 8 hours, about 0.5 hour to about 6 hours, about 0.5 hour to about 4 hours, about 0.5 hour to about 2 hours, about 0.5 Hour to about 1 hour, about 0.5 hour to about 1.5 hours, about 1 hour to about 24 hours, about 1 hour to about 20 hours, it is about 1 small Up to about 18 hours, about 1 hour to about 16 hours, about 1 hour to about 14 hours, about 1 hour to about 12 hours, about 1 hour extremely About 10 hours, about 1 hour to about 8 hours, about 1 hour to about 6 hours, about 1 hour to about 4 hours, about 1 hour to about 2 hours, About 2 hours to about 24 hours, about 2 hours to about 20 hours, about 2 hours to about 18 hours, about 2 hours to about 16 hours, it is about 2 small Up to about 14 hours, about 2 hours to about 12 hours, about 2 hours to about 10 hours, about 2 hours to about 8 hours, about 2 hours to about 6 hours, about 2 hours to about 4 hours, about 3 hours to about 24 hours, about 3 hours to about 20 hours, about 3 hours to about 18 hours, About 3 hours to about 16 hours, about 3 hours to about 14 hours, about 3 hours to about 12 hours, about 3 hours to about 10 hours, it is about 3 small Up to about 8 hours, about 3 hours to about 6 hours, about 3 hours to about 4 hours, about 4 hours to about 24 hours, about 4 hours to about 20 Hour, about 4 hours to about 18 hours, about 4 hours to about 16 hours, about 4 hours to about 14 hours, about 4 hours to about 12 hours, About 4 hours to about 10 hours, about 4 hours to about 8 hours, about 4 hours to about 6 hours, about 6 hours to about 24 hours, about 6 hours To about 20 hours, about 6 hours to about 18 hours, about 6 hours to about 16 hours, about 6 hours to about 14 hours, about 6 hours to about 12 hours, about 6 hours to about 10 hours, about 6 hours to about 8 hours, about 12 hours to about 24 hours, it is about 12 hours to about 20 small When, about 12 hours to about 18 hours, about 12 hours to about 16 hours, about 12 hours to about 14 hours, it is about 18 hours to about 24 small When between or be greater than 24 hours.For example, first time point can be between 0 hour to 6 hours, about 6 hours to 12 hours it Between, between about 12 hours to 18 hours or between about 18 hours to 24 hours.

In some embodiments, the second time point or optional third time point or the 4th time point are at the first time About 1 hour to about 10 hours after point, about 3 hours to about 6 hours after such as first time point.In some embodiments, Second time point was about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 after first time point Hour, about 8 hours, about 9 hours or about 10 hours.

In some embodiments, from the UCH-L1 level in the first sample that first time point obtains at second Between horizontal statistically significant of UCH-L1 in the second sample for obtaining of point or optional third time point or the 4th time point To increase or decrease be at least about 1 times, at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, At least about 6 times, at least about 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times or at least 20 times.In some embodiments, From the UCH-L1 level in the first sample that first time point obtains at the second time point or optional third time point or The statistically significant of UCH-L1 level in the second sample that 4th time point obtained is reduced by least about 1 times, at least about 1.5 Again, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least 8 times, at least 9 Times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, extremely Few 18 times, at least 19 times or at least 20 times show that subject will have positive head CT scan.In some embodiments, from The UCH-L1 level in the first sample that first time point obtains is at the second time point or optional third time point or the 4th At least about 1 times, at least about 1.5 times of the statistically significant increase, extremely for the UCH-L1 level in the second sample that time point obtains Few about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least 8 times, at least 9 times, extremely Few 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 Again, show that subject there will be positive head CT scan at least 19 times or at least 20 times.In some embodiments, from first The UCH-L1 level in the first sample that time point obtains is at the second time point or optional third time point or the 4th time Not statistically significant the increasing or decreasing of UCH-L1 level in the second sample that point obtains shows that subject will have yin Property head CT scan.

In some embodiments, from the UCH-L1 level in the second sample that the second time point obtained at first Between horizontal statistically significant of UCH-L1 in the first sample for obtaining of point or optional third time point or the 4th time point Increase or decrease be more than about 0.1 times, more than about 0.2 times, more than about 0.3 times, more than about 0.4 times, more than about 0.5 times, it is super Cross about 0.55 times, more than about 0.6 times, more than about 0.7 times, more than about 0.73 times, more than about 0.8 times, more than about 0.9 times, be more than About 1 times, more than 1.5 times, more than 1.81 times, more than about 2 times, more than about 3 times, more than about 4 times, more than about 5 times, more than about 6 times, More than about 7 times, more than 8 times, more than 9 times, more than 10 times, more than 11 times, more than 12 times, more than 13 times, more than 14 times, more than 15 Again, more than 16 times, more than 17 times, more than 18 times, more than 19 times or more than 20 times.In some embodiments, from second Between put UCH-L1 level in the second sample of acquirement at first time point or optional third time point or the 4th time point Horizontal statistically significant of the UCH-L1 in the first sample obtained reduce by more than about 0.1 times, more than about 0.2 times, be more than about 0.3 times, more than about 0.4 times, more than about 0.5 times, more than about 0.55 times, more than about 0.6 times, more than about 0.7 times, be more than about 0.73 Again, more than about 0.8 times, more than about 0.9 times, more than about 1 times, more than 1.5 times, more than 1.81 times, more than about 2 times, more than about 3 Times, more than about 4 times, more than about 5 times, more than about 6 times, more than about 7 times, more than 8 times, more than 9 times, more than 10 times, more than 11 times, More than 12 times, more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 times, more than 18 times, more than 19 times or be more than 20 times show that subject is likely to have positive head CT scan.In some embodiments, from obtaining at the second time point UCH-L1 level in second sample is to the first of first time point or the acquirement of optional third time point or the 4th time point Horizontal statistically significant of UCH-L1 in sample increase above about 0.1 times, more than about 0.2 times, more than about 0.3 times, be more than About 0.4 times, more than about 0.5 times, more than about 0.55 times, more than about 0.6 times, more than about 0.7 times, more than about 0.73 times, be more than about 0.8 times, more than about 0.9 times, more than about 1 times, be more than 1.5 times, be more than 1.81 times, more than about 2 times, be more than about 3 times, more than about 4 Times, more than about 5 times, more than about 6 times, more than about 7 times, more than 8 times, more than 9 times, more than 10 times, more than 11 times, more than 12 times, Show more than 13 times, more than 14 times, more than 15 times, more than 16 times, more than 17 times, more than 18 times, more than 19 times or more than 20 times Subject is likely to have positive head CT scan.In some embodiments, from the second sample obtained at the second time point In UCH-L1 level to first time point or optional third time point or the 4th time point acquirement the second sample in Not statistically significant the increasing or decreasing of UCH-L1 level shows that subject will likely have negative head CT scan.

In some embodiments, from the UCH-L1 level in the first sample that first time point obtains at second Between put statistically significant the increasing or decreasing of the UCH-L1 level in the second sample of acquirement and be less than about 0.1 times, be less than About 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, be less than about 1 times, less than 1.5 times, less than 1.81 times, it is small In about 2, less than about 3 times, less than about 4 times, less than about 5 times, less than about 6 times, less than about 7 times, less than 8 times, less than 9 times, be less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 times, less than 18 times, Less than 19 times or less than 20 times.In some embodiments, the UCH-L1 water from the first sample that first time point obtains Put down the second time point obtain the second sample in UCH-L1 level statistically significant reduction less than about 0.1 times, it is small In about 0.2 times, be less than about 0.3 times, be less than about 0.4 times, be less than about 0.5 times, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, be less than about 1 times, less than 1.5 times, less than 1.81 times, it is small In about 2 times, be less than about 3 times, be less than about 4 times, be less than about 5 times, be less than about 6 times, be less than about 7 times, be less than about 8 times, less than 9 times, Less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, less than 15 times, less than 16 times, less than 17 times, be less than 18 times, less than 19 times or less than 20 times show that subject will likely have positive CT scan.In some embodiments, Cong The UCH-L1 level in the first sample that one time point obtained is to the UCH-L1 water in the second sample that the second time point obtained Flat statistically significant increase is less than about 0.1 times, is less than about 0.2 times, is less than about 0.3 times, is less than about 0.4 times, is less than about 0.5 Again, at least about 0.55 times, at least about 0.6 times, at least about 0.7 times, at least about 0.73 times, at least about 0.8 times, at least about 0.9 times, Less than about 1 times, less than about 1.5 times, less than 1.81 times, less than about 2 times, less than about 3 times, less than about 4 times, less than about 5 times, be less than About 6 times, be less than about 7 times, less than 8 times, less than 9 times, less than 10 times, less than 11 times, less than 12 times, less than 13 times, less than 14 times, Show that subject there will likely be sun less than 15 times, less than 16 times, less than 17 times, less than 18 times, less than 19 times or less than 20 times Property head CT scan.In some embodiments, from the UCH-L1 level in the first sample that first time point obtains to Not statistically significant the increasing or decreasing of UCH-L1 level in the second sample that second time point obtained shows subject There will likely be negative head CT scan.

In some embodiments, when the horizontal of UCH-L1 decreases or increases at least absolutely from the first sample to the second sample When amount, subject is evaluated as to there may be positive head CT scan.In some embodiments, absolute magnitude is at least about 20pg/mL is between about 6100pg/mL.In some embodiments, absolute magnitude can be at least about 20pg/mL to about Between 6100pg/mL, at least about 20pg/mL between about 6000pg/mL, at least about 20pg/mL to about 5500pg/mL it Between, at least about 20pg/mL between about 5000pg/mL, at least about 20pg/mL between about 4500pg/mL, at least about 20pg/mL between about 4000pg/mL, at least about 20pg/mL between about 3500pg/mL, at least about 20pg/mL to about Between 3000pg/mL, at least about 20pg/mL between about 2500pg/mL, at least about 20pg/mL to about 2000pg/mL it Between, at least about 20pg/mL between about 1500pg/mL, at least about 20pg/mL between about 1000pg/mL, at least about 20pg/mL between about 900pg/mL, at least about 20pg/mL between about 800pg/mL, at least about 20pg/mL to about Between 700pg/mL, at least about 20pg/mL between about 600pg/mL, at least about 20pg/mL between about 500pg/mL, At least about 20pg/mL between about 400pg/mL, at least about 20pg/mL between about 300pg/mL, at least about 20pg/ ML between about 200pg/mL, at least about 20pg/mL between about 100pg/mL, at least about 20pg/mL to about 50pg/mL Between, at least about 25pg/mL between about 6100pg/mL, at least about 25pg/mL between about 6000pg/mL, at least About 25pg/mL between about 5500pg/mL, at least about 25pg/mL between about 5000pg/mL, at least about 25pg/mL extremely Between about 4500pg/mL, at least about 25pg/mL between about 4000pg/mL, at least about 25pg/mL to about 3500pg/mL Between, at least about 25pg/mL between about 3000pg/mL, at least about 25pg/mL between about 2500pg/mL, at least About 25pg/mL between about 2000pg/mL, at least about 25pg/mL between about 1500pg/mL, at least about 25pg/mL extremely Between about 1000pg/mL, at least about 25pg/mL between about 900pg/mL, at least about 25pg/mL to about 800pg/mL it Between, at least about 25pg/mL between about 700pg/mL, at least about 25pg/mL between about 600pg/mL, at least about 25pg/mL between about 500pg/mL, at least about 25pg/mL between about 400pg/mL, at least about 25pg/mL to about Between 300pg/mL, at least about 25pg/mL between about 200pg/mL, at least about 25pg/mL between about 100pg/mL, At least about 25pg/mL between about 50pg/mL, at least about 50pg/mL between about 6100pg/mL, at least about 50pg/ ML between about 6000pg/mL, at least about 50pg/mL between about 5500pg/mL, at least about 50pg/mL to about Between 5000pg/mL, at least about 50pg/mL between about 4500pg/mL, at least about 50pg/mL to about 4000pg/mL it Between, at least about 50pg/mL between about 3500pg/mL, at least about 50pg/mL between about 3000pg/mL, at least about 50pg/mL between about 2500pg/mL, at least about 50pg/mL between about 2000pg/mL, at least about 50pg/mL to about Between 1500pg/mL, at least about 50pg/mL between about 1000pg/mL, at least about 50pg/mL to about 900pg/mL it Between, at least about 50pg/mL between about 800pg/mL, at least about 50pg/mL between about 700pg/mL, at least about 50pg/mL between about 600pg/mL, at least about 50pg/mL between about 500pg/mL, at least about 50pg/mL to about Between 400pg/mL, at least about 50pg/mL between about 300pg/mL, at least about 50pg/mL between about 200pg/mL, At least about 50pg/mL between about 100pg/mL, at least about 100pg/mL between about 6100pg/mL, at least about 100pg/mL between about 6000pg/mL, at least about 100pg/mL between about 5500pg/mL, at least about 100pg/mL To between about 5000pg/mL, at least about 100pg/mL between about 4500pg/mL, at least about 100pg/mL to about Between 4000pg/mL, at least about 100pg/mL between about 3500pg/mL, at least about 100pg/mL to about 3000pg/mL Between, at least about 100pg/mL between about 2500pg/mL, at least about 100pg/mL between about 2000pg/mL, extremely Few about 100pg/mL between about 1500pg/mL, at least about 100pg/mL between about 1000pg/mL, at least about 100pg/mL between about 900pg/mL, at least about 100pg/mL between about 800pg/mL, at least about 100pg/mL extremely Between about 700pg/mL, at least about 100pg/mL between about 600pg/mL, at least about 100pg/mL to about 500pg/mL Between, at least about 100pg/mL between about 400pg/mL, at least about 100pg/mL between about 300pg/mL, at least About 100pg/mL between about 200pg/mL, at least about 129pg/mL between about 6100pg/mL, at least about 129pg/mL To between about 6000pg/mL, at least about 129pg/mL between about 5500pg/mL, at least about 129pg/mL to about Between 5000pg/mL, at least about 129pg/mL between about 4500pg/mL, at least about 129pg/mL to about 4000pg/mL Between, at least about 129pg/mL between about 3500pg/mL, at least about 129pg/mL between about 3000pg/mL, extremely Few about 129pg/mL between about 2500pg/mL, at least about 129pg/mL between about 2000pg/mL, at least about 129pg/mL between about 1500pg/mL, at least about 129pg/mL between about 1000pg/mL, at least about 129pg/mL To between about 900pg/mL, at least about 129pg/mL between about 800pg/mL, at least about 129pg/mL to about 700pg/ Between mL, at least about 129pg/mL between about 600pg/mL, at least about 129pg/mL between about 500pg/mL, extremely Few about 129pg/mL between about 400pg/mL, at least about 129pg/mL between about 300pg/mL, at least about 129pg/ ML between about 200pg/mL, at least about 200pg/mL between about 6100pg/mL, at least about 200pg/mL to about Between 6000pg/mL, at least about 200pg/mL between about 5500pg/mL, at least about 200pg/mL to about 5000pg/mL Between, at least about 200pg/mL between about 4500pg/mL, at least about 200pg/mL between about 4000pg/mL, extremely Few about 200pg/mL between about 3500pg/mL, at least about 200pg/mL between about 3000pg/mL, at least about 200pg/mL between about 2500pg/mL, at least about 200pg/mL between about 2000pg/mL, at least about 200pg/mL To between about 1500pg/mL, at least about 200pg/mL between about 1000pg/mL, at least about 200pg/mL to about Between 900pg/mL, at least about 200pg/mL between about 800pg/mL, at least about 200pg/mL to about 700pg/mL it Between, at least about 200pg/mL between about 600pg/mL, at least about 200pg/mL between about 500pg/mL, at least about 200pg/mL between about 400pg/mL, at least about 200pg/mL between about 300pg/mL, at least about 300pg/mL extremely Between about 6100pg/mL, at least about 300pg/mL between about 6000pg/mL, at least about 300pg/mL to about 5500pg/ Between mL, at least about 300pg/mL between about 5000pg/mL, at least about 300pg/mL between about 4500pg/mL, In At least about 300pg/mL between about 4000pg/mL, at least about 300pg/mL between about 3500pg/mL, at least about 300pg/mL between about 3000pg/mL, at least about 300pg/mL between about 2500pg/mL, at least about 300pg/mL To between about 2000pg/mL, at least about 300pg/mL between about 1500pg/mL, at least about 300pg/mL to about Between 1000pg/mL, at least about 300pg/mL between about 900pg/mL, at least about 300pg/mL to about 800pg/mL it Between, at least about 300pg/mL between about 700pg/mL, at least about 300pg/mL between about 600pg/mL, at least about 300pg/mL between about 500pg/mL, at least about 300pg/mL between about 400pg/mL, at least about 400pg/mL extremely Between about 6100pg/mL, at least about 400pg/mL between about 6000pg/mL, at least about 400pg/mL to about 5500pg/ Between mL, at least about 400pg/mL between about 5000pg/mL, at least about 400pg/mL between about 4500pg/mL, In At least about 400pg/mL between about 4000pg/mL, at least about 400pg/mL between about 3500pg/mL, at least about 400pg/mL between about 3000pg/mL, at least about 400pg/mL between about 2500pg/mL, at least about 400pg/mL To between about 2000pg/mL, at least about 400pg/mL between about 1500pg/mL, at least about 400pg/mL to about Between 1000pg/mL, at least about 400pg/mL between about 900pg/mL, at least about 400pg/mL to about 800pg/mL it Between, at least about 400pg/mL between about 700pg/mL, at least about 400pg/mL between about 600pg/mL, at least about 400pg/mL between about 500pg/mL, at least about 500pg/mL between about 6100pg/mL, at least about 500pg/mL extremely Between about 6000pg/mL, at least about 500pg/mL between about 5500pg/mL, at least about 500pg/mL to about 5000pg/ Between mL, at least about 500pg/mL between about 4500pg/mL, at least about 500pg/mL between about 4000pg/mL, In At least about 500pg/mL between about 3500pg/mL, at least about 500pg/mL between about 3000pg/mL, at least about 500pg/mL between about 2500pg/mL, at least about 500pg/mL between about 2000pg/mL, at least about 500pg/mL To between about 1500pg/mL, at least about 500pg/mL between about 1000pg/mL, at least about 500pg/mL to about Between 900pg/mL, at least about 500pg/mL between about 800pg/mL, at least about 500pg/mL to about 700pg/mL it Between or at least about 500pg/mL between about 600pg/mL.In some embodiments, absolute magnitude can be at least about 20pg/ ML, at least about 21pg/mL, at least about 22pg/mL, at least about 23pg/mL, at least about 24pg/mL, at least about 25pg/mL, at least About 26pg/mL, at least about 27pg/mL, at least about 28pg/mL, at least about 29pg/mL, at least about 30pg/mL, at least about 35pg/ ML, at least about 40pg/mL, at least about 45pg/mL, at least about 50pg/mL, at least about 55pg/mL, at least about 60pg/mL, at least About 65pg/mL, at least about 70pg/mL, at least about 75pg/mL, at least about 80pg/mL, at least about 85pg/mL, at least about 90pg/ ML, at least about 95pg/mL, at least about 100pg/mL, at least about 110pg/mL, at least about 120pg/mL, at least about 129pg/mL, At least about 130pg/mL, at least about 140pg/mL, at least about 150pg/mL, at least about 200pg/mL, at least about 250pg/mL, extremely Few about 300pg/mL, at least about 350pg/mL, at least about 400pg/mL, at least about 450pg/mL, at least about 500pg/mL, at least About 550pg/mL, at least about 600pg/mL, at least about 700pg/mL, at least about 800pg/mL, at least about 900pg/mL, at least about 1000pg/mL, at least about 1500pg/mL, at least about 2000pg/mL, at least about 2500pg/mL, at least about 2528pg/mL, extremely Few about 3000pg/mL, at least about 4000pg/mL, at least about 5000pg/mL, at least about 6000pg/mL or at least about 6100pg/ mL。

6, early detection is helped to suffer from the slight or moderate in the human experimenter of the damage to head to severe trauma The method of property cerebral injury

This disclosure relates to which a kind of help early detection suffers from or may suffer from the human experimenter of the damage to head In traumatic brain injury method.The method can help early detection to suffer from or may suffer from the damage to head Human experimenter in traumatic brain injury (such as determine subject whether suffer from traumatic brain injury).As used herein, " determining whether subject suffers from traumatic brain injury " refer to using the method (such as using such as clinical assessment data its Its information) determine that subject is more likely to TBI.The method includes to the sample from human experimenter be measured with The level of measurement or detection the first sample and the ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in the second sample.First sample exists To being derived from human experimenter in after head injury 24 hours and the second sample is about 3 hours to about 6 small after obtaining the first sample When after be derived from human experimenter.Determine that UCH-L1's is horizontal from the first sample to the reduction of the second sample or decline, and if There is statistically significant reduction from the UCH-L1 level in the first sample to the UCH-L1 level in the second sample, then it will be by Examination person is evaluated as with slight or moderate to severe trauma cerebral injury.Sample can be biological sample.

In some embodiments, statistically significant reduction includes lower at least than the UCH-L1 level in the first sample 5% UCH-L1 level reduces or decline.In some embodiments, statistically significant reduction includes that UCH-L1 level subtracts Less or decline, wherein the UCH-L1 level in the second sample is horizontal lower by about 5% to about 1000% than the UCH-L1 in the first sample. For example, UCH-L1 level can more horizontal than UCH-L1 in the first sample low at least about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, About 900% or about 1000%.In some embodiments, UCH-L1 level can than the UCH-L1 level in the first sample down to Few about 1 times, at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 Times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, at least 16 times, at least 17 times, at least 18 times, at least 19 times or at least 20 times.

In some embodiments, statistically significant reduction includes the reduction or decline of UCH-L1 level, wherein the UCH-L1 level in two samples is low at least about 20pg/mL at least about 6100pg/mL more horizontal than the UCH-L1 in the first sample.

Optionally, the method further includes will such as in step (b) measure UCH-L1 concentration or amount for example with ginseng The level of examining is compared.In addition, if the relatively display such as concentration of the UCH-L1 of measurement in step (b) or amount is for example opposite In the unfavorable change of reference levels, then the method, which is optionally included, continues one with one or more medicine composite for curing subjects The section time.

Specifically, relative to for monitoring progression of disease and/or treatment or for determining that subject develops slight or moderate To the reference levels of the risk of severe trauma cerebral injury, the amount or concentration of UCH-L1 or UCH-L1 segment, which can be, " not to be changed ", " advantageous " (or " favorably changing ") or " unfavorable " (or " unfavorable change ")." raising " or " increase " refers to test Amount or concentration in sample are higher than or greater than typical or normal level or range (such as reference levels) or are higher than or greater than another Reference levels or range (such as previous or baseline sample).Term " decline " or " reduction " refer to amount or concentration in test sample Below or less than typical or normal level or range (such as reference levels) or it is below or less than another reference levels or range (example Such as previous or baseline sample).Term " change " refers to that amount in sample or concentration and typical or normal level or range (such as are joined Examine level) it compares or changes that (increasing adds deduct compared with another reference levels or range (such as previously or baseline sample) It is few).

The typical case of UCH-L1 or normal level or range are defined according to standard practices.When with typical or normal level Or range or reference levels or range are compared, when there is any net change that cannot be explained by experimental error or sample variation, It can be considered the level or variation of so-called change occurred.Therefore, by measured in specific sample level with from it is so-called just The level or horizontal extent measured in the similar sample of normal subject compares.In that case, " normal subjects " are not Individual with detectable disease or illness, and " normal " (sometimes referred to as " compares ") patient or group is for example non-table respectively Reveal patient or the group of detectable disease or illness." apparent normal subjects " are that UCH-L1 is not yet evaluated or assessed Subject (i.e. not with the individual of detectable disease or illness).Arrived when analyte is usually undetectable (such as normal level It is zero, or in the range of about the 25 of normal population to about 75 percentile), but when being detected in the test sample, and When analyte under being higher than normal level when being present in test sample, then it is assumed that the level " raising " of analyte.Therefore, this public affairs It opens and especially provides a kind of screening with slight or moderate to severe trauma cerebral injury or in slight or moderate to severe The method of subject in the risk of traumatic brain injury.

Reference levels in this method can be with slight or moderate into the patient of severe trauma cerebral injury UCH-L1 is horizontal.In some embodiments, in serum be greater than or equal to 5pg/mL, 10pg/mL, 20pg/mL, 30pg/mL, 40pg/mL、50pg/mL、60pg/mL、70pg/mL、80pg/mL、90pg/mL、100pg/mL、500pg/mL、1000pg/mL、 Subject is accredited as with slight or moderate to weight by the UCH-L1 level of 5000pg/mL, 10000pg/mL or 50000pg/mL Spend traumatic brain injury.Optionally, in some cases, in serum be greater than or equal to 100000pg/mL, 500000pg/mL, Subject is accredited as trouble by the UCH-L1 level of 1000000pg/mL, 150000pg/mL, 200000pg/mL or 500000pg/mL There is slight or moderate to severe trauma cerebral injury.

In some embodiments, the method further includes being confirmed as carrying out with the treatment of traumatic brain injury therapy The human experimenter of CT scan, as described below.In some embodiments, the method further includes as described below Monitoring is confirmed as carrying out the human experimenter of CT scan.

7, the combination of UCH-L1 and other biomarkers

In some embodiments, a kind of or more to measure or detect the method further includes being measured to sample The level of the other biomarkers of kind.The measurement can be for one of measurement or sample or a variety of other biomarkers The horizontal measuring method of object, immunoassay, mass spectrum etc..In addition, measuring method can be used in the form of clinical chemistry, such as It will be by known to persons of ordinary skill in the art.In some embodiments, the measurement includes: the first sample of measurement or detection With the level of one of the second sample or a variety of other biomarkers, determine the water of one or more other biomarkers It puts down from the first sample to the increase of the second sample and/or reduction or rises or falls, and if from one of first sample Or a variety of other biomarker levels have statistics to one of the second sample or a variety of other biomarker levels On significantly increase or decrease, then human experimenter is evaluated as with slight or moderate to severe trauma cerebral injury.

It is contemplated that compared with measuring individual UCH-L1, UCH-L1 and the one or more biologies special to disease The combination of marker or immunoassay can provide bigger discrimination between normal healthy controls and individual with disease.Example Such as, compared with individual UCH-L1 group, the group for measuring UCH-L1 and other traumatic brain injury biomarker can be strong Bigger discrimination is provided between health control and the individual with disease.UCH-L1 and at least one or more of biomarker Combination can provide bigger discrimination between normal healthy controls and individual with slight or moderate to severe trauma cerebral injury Power.The example of one or more biomarkers includes glial fibrillary acidic protein (GFAP), S100 calbindin B (S100 β), neuron specific enolase (NSE), crystallin B chain (CRYAB), brain lipid binding protein (BLBP), aldehyde Contracting enzyme C (ALDOC), astroglia phosphoprotein 15 (PEA15), glutamine synthelase (GS), Apo lipoprotein 1, Tau, C Reactive protein (CRP), free brain-derived neurotrophic factor (BDNF), p-Tau, total BDNF, Troponin I (TnI) and its group It closes.In some embodiments, one or more other biomarkers are S100 β, CRP, Apo lipoprotein 1 or crystalline lens egg White B chain (CRYAB).

8, to the treatment and monitoring of the subject for suffering from traumatic brain injury

It can treat or monitor and be accredited or be evaluated as in the above-mentioned methods with traumatic brain injury, such as mild trauma Property cerebral injury or moderate to severe trauma cerebral injury subject.In some embodiments, the method further includes The people with traumatic brain injury is assessed as with traumatic brain injury therapy (all any therapies as known in the art) treatment Class subject.For example, the treatment of traumatic brain injury can take many forms, this depends on the serious of the damage to head Property.For example, treatment may include rest, avoid body movement (such as moving), avoid for the subject for suffering from slight TBI Light one of wears sunglasses, the drug for alleviating headache or migraine, anti-nausea drug etc. or a variety of when outgoing. The treatment of the patient of suffering from serious TBI may include and give one or more drugs appropriate (such as diuretics, anticonvulsion Drug has been placed in drug in drug-induced stupor or other pharmacy or bio-pharmaceuticals drug (for calmness and by individual Know for or the following exploitation for treating TBI's), one or more surgical operations (such as removal hemotoncus, repair skull bone Folding, decompression craniectomy etc.) and one or more therapies (such as one or many rehabilitations, cognitive-behavioral therapy, indignation pipe Reason, Counseling Psychology etc.).In some embodiments, the method further includes monitorings to be assessed as with traumatic brain The human experimenter of damage (such as slight or moderate to severe trauma).In some embodiments, can with CT scan or MRI monitoring is accredited as with traumatic brain injury, such as mild trauma cerebral injury or severe trauma cerebral injury it is tested Person.

9, for measuring the horizontal method of UCH-L1

In method as discussed above, UCH-L1 level can be measured by any means, the means such as antibody according to Rely property method, such as immunoassay, the Western Immuno precipitation method, immunoelectrophoresis, chemical analysis, SDS-PAGE and albumen Matter engram analysis or Western Immuno decoration method, electrophoretic analysis, protein determination, competitive binding assay, functionality Protein determination or chromatography or spectroscopic methodology, such as high performance liquid chromatography (HPLC) or liquid chromatography-mass spectrography (LC/MS). In addition, measuring method can be used in the form of clinical chemistry, it such as will be by known to persons of ordinary skill in the art.

In some embodiments, the level for measuring UCH-L1 includes making sample and the first specific binding members and second Specific binding members contact.In some embodiments, the first specific binding members are capture antibody, and second is special Property binding members be detection antibody.In some embodiments, the level for measuring UCH-L1 includes making sample simultaneously or with any Sequence is successively contacted with following: (1) capturing antibody, it is anti-to form capture in conjunction with the epitope in UCH-L1 or UCH-L1 segment Body-UCH-L1 antigenic compound, and (2) detect antibody, it includes detectable label and combine UCH-L1 on it is at large obtain it is anti- The epitope that body combines, to form UCH-L1 antigen-detection antibody complex, so that forming capture antibody-UCH-L1 antigen-detection Antibody complex, and based on by capturing the detectable label generation in antibody-UCH-L1 antigen-detection antibody complex Signal measures the amount or concentration of UCH-L1 in sample.

In some embodiments, the first specific binding members are immobilized on solid support.In some implementations In mode, the second specific binding members are immobilized on solid support.In some embodiments, the first specificity knot The UCH-L1 antibody that synthesis person is discussed further below.

In some embodiments, sample is diluted or not diluted.Sample can be about 1 to about 25 microlitre, About 1 to about 24 microlitre, about 1 to about 23 microlitre, about 1 to about 22 microlitre, about 1 to about 21 microlitre, about 1 to about 20 microlitre, about 1 to about 18 microlitres, about 1 to about 17 microlitre, about 1 to about 16 microlitre, about 15 microlitres or about 1 liter, about 2 microlitres, about 3 microlitres, about 4 microlitres, about 5 microlitres, about 6 microlitres, about 7 microlitres, about 8 microlitres, about 9 microlitres, about 10 microlitres, about 11 microlitres, about 12 microlitres, about 13 microlitres, about 14 microlitres, about 15 microlitres, about 16 microlitres, about 17 microlitres, about 18 microlitres, about 19 microlitres, about 20 microlitres, about 21 microlitres, it is about 22 micro- It rises, about 23 microlitres, about 24 microlitres or about 25 microlitres.In some embodiments, sample be about 1 to about 150 microlitre or less or About 1 to about 25 microlitre or less.

Some instruments in addition to point-of-care device are (as Abbott Laboratories instrument With other core laboratory instruments) it can measure the UCH-L1 level for being higher than or greater than 25000pg/mL in sample.

Other detection methods include that using nanopore device or nanometer well device or may be adapted in nanopore device or nanometer It is used on well device.The example of nanopore device is described in International Patent Publication No. WO 2016/161402, accordingly to draw Mode is integrally incorporated.The example of nanometer well device is described in International Patent Publication No. WO 2016/161400, accordingly It is incorporated hereby

10, UCH-L1 antibody

Specific binding ubiquitin carboxy terminal hydrolase-l 1 (" UCH-L1 ") (or its piece can be used in method described herein Section) isolated antibody, referred to as " UCH-L1 antibody ".UCH-L1 antibody can be used for assessing the measurement as traumatic brain injury UCH-L1 state, the presence of UCH-L1 in test sample, UCH-L1 present in quantitative sample amount or test sample in The presence of UCH-L1 and its quantitative amount.As used in the entire disclosure, term " measurement ", " detection " and " quantitative " and its appoint What derivative is used to describe the method disclosed herein for assessing UCH-L1 level in sample in which can be interchanged.

A, ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1)

Ubiquitin carboxy terminal esterase L1 (" UCH-L1 ") is also referred to as " ubiquitin c-terminal hydrolase " deubiquitinating enzymes. UCH-L1 is the small C-terminal adduct of its product hydrolysis ubiquitin to generate the member of the gene family of ubiquitin monomer.UCH-L1's Expressing has high degree of specificity to the cell of neuron and diffusivity neuroendocrine system and its tumour.It is largely present in institute Have in neuron (1%-2% of the total brain albumen of Zhan), it is specific expressed in neuron and testis/ovary.The catalysis of UCH-L1 Triplet contain the 90th cysteine, in the 176th aspartic acid and in the 161st histidine, they are responsible for Its hydrolytic enzyme activities.

People UCH-L1 can have following amino acid sequence:

MQLKPMEINPEMLNKVLSRLGVAGQWRFVDVLGLEEESLGSVPAPACALLLLFPLTAQHENFRKKQIE ELKGQEVSPKVYFMKQTIGNSCGTIGLIHAVANNQDKLGFEDGSVLKQFLSETEKMSPEDRAKCFEKNEAIQAAHD AVAQEGQCRVDDKVNFHFILFNNVDGHLYELDGRMPFPVNHGASSEDTLLKDAAKVCREFTEREQGEVRFSAVALC KAA (SEQ ID NO:1).

People UCH-L1 can be the segment or variant of SEQ ID NO:1.The length of the segment of UCH-L1 can be 5 with Between 225 amino acid, between 10 and 225 amino acid, between 50 and 225 amino acid, between 60 and 225 amino acid, Between 65 and 225 amino acid, between 100 and 225 amino acid, between 150 and 225 amino acid, 100 and 175 amino Between acid or between 175 and 225 amino acid.Segment may include multiple continuous amino acids from SEQ ID NO:1.

B, UCH-L1 identifies antibody

Antibody is the antibody in conjunction with UCH-L1, its segment, the epitope of UCH-L1 or its variant.Antibody can be anti-UCH- The segment of L1 antibody or its variant or derivative.Antibody can be polyclonal antibody or monoclonal antibody.Antibody can be chimeric Antibody, single-chain antibody, affinity maturation antibody, human antibody, humanized antibody, fully human antibodies or antibody fragment (such as Fab piece Or mixtures thereof section),.Antibody fragment or derivative may include F (ab')₂, Fv or scFv segment.Antibody derivatives can pass through Simulating peptide generates.In addition, description can be adapted for generating single-chain antibody for generating the technology of single-chain antibody.

Anti- UCH-L1 antibody can be inosculating antibody UCH-L1 or the anti-UCH-L1 antibody of humanization.In one embodiment, Humanized antibody and chimeric antibody are all monovalent.In one embodiment, humanized antibody and chimeric antibody include with The single area Fab of the area Fc connection.

Human antibody can be originated from display technique of bacteriophage or the transgenic mice from expression human immunoglobulin gene.People The result that antibody can be used as immune response in human body is generated and is separated.See, for example, Funaro et al., BMC Biotechnology,2008(8):85.Therefore, antibody can be the product of people and non-animal pedigree.Because it is humanized , it is possible to minimize the risk reacted autoantigen.Optionally, test yeast display libraries and display technique can be used for Selection and the separation anti-UCH-L1 antibody of people.For example, the library of natural human single chain variable fragment (scFv) can be used for selecting people anti- UCH-L1 antibody.Transgenic animals can be used for expressing human antibody.

Humanized antibody can be the antibody from non-human species in conjunction with required antigen, come from one or more The complementary determining region (CDR) of non-human species and the framework region from human immunoglobulin molecule.

The antibody is that it has the biology function different with antibody as known in the art from the difference of known antibodies Energy.

(1) epitope

Antibody can be with immunologic specificity combination UCH-L1 (SEQ ID NO:1), its segment or its variant.Antibody can be immunized Specific recognition simultaneously combines at least three amino acid, at least four amino acid, at least five amino acid, at least in epitope regions Six amino acid, at least seven amino acid, at least eight amino acid, at least nine amino acid or at least ten amino acid.Antibody Can identify and combine such epitope with immunologic specificity, the epitope have epitope regions at least three continuous amino acids, At least four continuous amino acids, at least five continuous amino acids, at least six continuous amino acids, at least seven continuous amino acids, At least eight continuous amino acids, at least nine continuous amino acids or at least ten continuous amino acids.

C, Antibody preparation/generation

Antibody can be prepared by any one of multiple technologies, including those technologies well known to those skilled in the art. In general, antibody can be generated by cell culture technology, and the cell culture technology includes via routine techniques or passing through Antibody gene, heavy chain and/or light chain are transfected into suitable bacterium or mammalian cell host to allow to generate antibody (its Middle antibody can be recombination) generation of Lai Jinhang monoclonal antibody.Various forms of terms " transfection " are intended to cover usually use In the extensive multiple technologies introducing exogenous DNA in protokaryon or eukaryotic host cell, for example, electroporation, calcium phosphate precipitation, DEAE- polydextrose transfection etc..Although antibody can be expressed in protokaryon or eukaryotic host cell, preferably in eukaryocyte and Most preferably express antibody in mammalian host cell because such eukaryocyte (and especially mammalian cell) compared to Prokaryotic cell is more likely to assemble and secrete the correct antibody folded and have immunologic competence.

Exemplary mammals host cell for expressing recombinant antibody includes Chinese hamster ovary cell (Chinese hamster ovary celI) (including Urlaub and Chasin, Proc.Natl.Acad.Sci.USA, dhfr-CHO described in 77:4216-4220 (1980) Cell), it is used together with DHFR selectable marker, such as such as Kaufman and Sharp, J.Mol.Biol, 159:601- Described in 621 (1982)；NS0 myeloma cell；COS cell and SP2 cell.When by the recombinant expression carrier of encoding antibody genes When being introduced into mammalian host cell, by by host cell culture be enough to allow antibody expressed in host cell or The a period of time in culture medium for more preferably growing antibody-secreting to host cell generates antibody.Mark can be used in antibody Quasi- method of purifying protein is recycled from culture medium.

Host cell can also be used for generating functional antibody fragment, such as Fab segment or scFv molecule.It should be appreciated that can To carry out modification to above procedure.For example, it may be possible to it is desirable that with the light chain of encoding antibody and/or the functional piece of heavy chain The DNA transfection host cell of section.Recombinant DNA technology can also be used for removing some or all codings and non-binding antigen institute interested The DNA of any of required light chain and heavy chain or both.The antibody is also covered by point that DNA molecular expression is truncated by this class Son.In addition, bifunctional antibody can be generated by being crosslinked antibody and secondary antibody with standard chemical cross-linking method, one of them Heavy chain and a light chain are that antibody (combining people UCH-L1) and another heavy chain and another light chain have the antigen of inhuman UCH-L1 There is specificity.

In an optimum decision system for recombinantly expressing antibody or its antigen-binding portion thereof, pass through calcium phosphate mediation The recombinant expression carrier of both encoding antibody heavy and antibody light chain is introduced into dhfr-CHO cell by infection protocol.In recombination table Up in carrier, heavy chain of antibody and light chain gene is made to be each operably linked to cmv enhancer/AdMLP promoter regulation element To drive gene height to transcribe.Recombinant expression carrier allows to select using methotrexate (MTX) selection/amplification with DHFR gene Select the Chinese hamster ovary celI transfected with carrier.Selected transformant host cell is cultivated to allow to express heavy chain of antibody and light chain And complete antibody is recycled from culture medium.Standard molecular biological technique is used to prepare recombinant expression carrier, transfecting host Cell, selection transformant, culture host cell, and antibody is recycled from culture medium.Further, recombinant antibodies are synthesized Method can be by cultivating host cell in being suitble to culture medium, until synthesis recombinant antibodies carry out.The method can be into one Step includes that recombinant antibodies are separated from culture medium.

The method of preparation monoclonal antibody includes the immortal cell line that preparation can generate the antibody with required specificity. Such cell line can be generated by the splenocyte obtained from immune animal.It can be immune with UCH-L1 or its segment and/or variant Animal.Peptide for animal to be immunized may include the amino of the tail region of encoding human Fc (such as crystallizable district fragment) or human antibody Acid.May then pass through for example makes immortalizing spleen cells with merging for myeloma cell fusion partner.It can be melted using a variety of Conjunction technology.For example, splenocyte and myeloma cell can be merged a few minutes with nonionic detergent, then it is inoculated with low-density On the selective medium supported hybrid cell growth but do not support myeloma cell growth.Technology as a kind of uses secondary Xanthine, aminopterin-induced syndrome, thymidine (HAT) selection.Another technology includes electro' asion.By time enough, normally about 1 to 2 Zhou Hou observes the colony of hybrid.It selects single colony and tests its culture supernatants to the combination activity of polypeptide.It can be with Use the hybridoma with high response and specificity.

Monoclonal antibody can be separated from the supernatant of the hybridoma colonies of growth.Furthermore, it is possible to using various technologies It improves yield, is such as injected into hybridoma cell line in the cavum peritoneale of suitable vertebrate host (such as mouse).So After can harvest monoclonal antibody from ascites or blood.Routine techniques, such as chromatography, gel filtration, precipitating can be passed through Pollutant is removed from antibody with extracting.Affinity chromatography is the example for the method that can be used during antibody purification.

Proteolytic enzyme papain cracks IgG molecule preferentially to generate several segments, two of them (F (ab) segment) Respectively contain the covalent heterodimer containing complete antigen binding site.It is several to provide that pepsin can crack IgG molecule Segment, including the F (ab') comprising two antigen binding sites₂Segment.

Fv segment can be by the preferential proteolytic hydrolytic rupture of IgM and ball is immunized in proteolytic cleavage IgG or IgA once in a while Protein molecular generates.The derivative Fv segment of recombinant technique can be used.Fv segment includes non-covalent VH::VL heterodimer, packet Containing remaining most of antigen binding ability of natural antibody mole-cules and the antigen binding site of binding ability.

Antibody, antibody fragment or derivative may include being plugged on heavy chain framework (" FR ") group and light chain framework (" FR ") respectively Complementary determining region of heavy chain (" CDR ") group and complementary determining region of light chain (" CDR ") group, the frame set between group provide for CDR It supports and limits the spatial relationship of CDR relative to each other.Three hypervariable regions of the CDR group containing heavy chain or the area light chain V.

Generation or separation, which can be used, has other appropriate methods of required specific antibody, including but not limited to from peptide Or selection in protein library (such as, but not limited to bacteriophage, ribosomes, oligonucleotides, RNA, cDNA, yeast etc. show library) The method of recombinant antibodies；For example, as can be from various commercial suppliers such as Cambridge using method as known in the art Antibody Technologies(Cambridgeshire,UK)、MorphoSys(Martinsreid/Planegg,Del.)、 What Bi ovation (Aberdeen, Scotland, UK) BioInvent (Lund, Sweden) was obtained.Referring to U.S. Patent number 4, 704,692,5,723,323,5,763,192,5,814,476,5,817,483,5,824,514,5,976,862.Alternative side Method is dependent on the immune transgenic mice that can generate human antibody pedigree (for example, SCID mice, Nguyen et al. (1997) Microbiol.Immunol.41:901-907；Sandhu et al. (1996) Crit.Rev.Biotechnol.16:95-118； Eren et al. (1998) Immunol.93:154-161), as known in the art and/or as described herein.Such technology Including but not limited to ribosomal display (Hanes et al. (1997) Proc.Natl.Acad.Sci.USA, 94:4937-4942； Hanes et al. (1998) Proc.Natl.Acad.Sci.USA, 95:14130-14135)；Monoclonal antibody generation technology (example Such as, lymphocyte antibody method (" SLAM ") (U.S. Patent number 5,627,052, Wen et al. (1987) of selection J.Immunol.17:887-892；Babcook et al. (1996) Proc.Natl.Acad Sci.USA 93:7843-7848)；It is solidifying Glue droplet and flow cytometry (Powell et al. (1990) Biotechnol.8:333-337；One Cell Systems, (Cambridge,Mass).；Gray et al. (1995) J.Imm.Meth.182:155-163；Kenny et al. (1995) Bio/ Technol.13:787-790)；B cell selection (Steenbakkers et al. (1994) Molec.Biol.Reports 19: 125-134(1994))。

Affinity maturation antibody can be generated by any one of multiple programs as known in the art.For example, with reference to Marks et al., BioTechnology 10:779-783 (1992) are described through the affine of VH and VL structural domain reorganization progress Power is mature.To the random mutagenesis of CDR and/or Framework residues by being described below: Barbas et al., Proc.Nat.Acad.Sci.USA,91:3809-3813(1994)；Schier et al., Gene, 169:147-155 (1995)； Yelton et al., J.Immunol., 155:1994-2004 (1995)；Jackson et al., J.Immunol., 154 (7): 3310- 3319(1995)；Hawkins et al., J.Mol.Biol, 226:889-896 (1992).In selective mutagenesis position and contacting Or super mutated site is described in 6,914,128 B1 of U.S. Patent number by the selective mutation that increased activity amino acid residue carries out.

Antibody variants can also be prepared using following manner: by the delivery of polynucleotides of encoding antibody to suitable host, To provide the transgenic animals or mammal that generate in its milk such antibody, goat, ox, horse, sheep etc. are made It is standby.These methods are well known in the art and are for example described in U.S. Patent number 5,827,690,5,849,992,4, 873,316, in 5,849,992,5,994,616,5,565,362 and 5,304,489.

Antibody variants can also be prepared in the following manner: delivery of polynucleotides is to provide the plant of genetically modified plants and culture Cell (such as, but not limited to tobacco, corn and duckweed), they generate such anti-in plant part or the cell cultivated by it Body, specific part or variant.For example, Cramer et al. (1999) Curr.Top.Microbiol.Immunol 240:95-118 It is described with references cited therein and for example generates the transgenosis cigarette for expressing a large amount of recombinant proteins using inducible promoter Blade of grass.Transgenic corns have been used to commercial production level express mammalian proteins, bioactivity with other heavy It is being generated in group system or suitable from those of natural origin purifying.See, for example, Hood et al., Adv.Exp.Med.Biol. (1999) 464:127-147 and references cited therein.Antibody variants can also be such as single-stranded anti-by comprising antibody fragment The transgenic plant seed of body (scFv), including tobacco seed and potato tubers largely generate.See, e.g. Conrad et al. (1998) Plant Mol.Biol.38:101-109 and references cited therein.It therefore, can also be with according to known method Antibody is generated using genetically modified plants.

It combines for example, immunogenicity or reduction, enhancing can be modified by addition exogenous sequence or modified, is affine Power, association rate, dissociation rate, affinity, specificity, half-life period or any other suitable feature are derivative to generate antibody Object.In general, keep it is inhuman or people's CDR sequence part or all, while employment or other amino acid substitution variable regions With the nonhuman sequence of constant region.

Small antibody fragment can be tool there are two the binary of antigen binding site, and wherein segment is contained in same polypeptide chain The heavy-chain variable domains (VH) being connect in (VH VL) with light-chain variable domain (VL).See, for example, EP 404,097；WO 93/ 11161；With Hollinger et al., (1993) Proc.Natl.Acad.Sci.USA90:6444-6448.By using too short Do not allow the connector matched between two structural domains in same chain, the complementary domain of structural domain and another chain is forced to match And generate two antigen binding sites.See also the U.S. Patent number 6 for authorizing Chen et al., 632,926, the patent is to quote Mode be integrally incorporated herein, and disclose antibody variants, it is anti-that there is the antibody variants one or more to be inserted into parent Amino acid in the hypervariable region of body and to the binding affinity of target antigen than parental antibody to the binding affinity of antigen by force extremely It is about 2 times few.

Antibody can be linear antibodies.The method for being used to prepare linear antibodies is well known in the art and is described in Zapata et al., (1995) Protein Eng.8 (10): 1057-1062.In brief, these antibody include a pair of series Fd Segment (VH-CH1-VH-CH1), the segment form a pair of of antigen binding domain.Linear antibodies can be bispecific or Dan Te Anisotropic.

It can be recycled from recombinant cell culture thing by known method and antibody purification, the known method includes but not It is limited to Protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation-exchange chromatography, phosphocellulose chromatography Method, hydrophobic interaction chromatography, affinity chromatography, hydroxyapatite chromatography and agglutinin chromatography.High performance liquid chromatography Method (" HPLC ") can also be used for purifying.

Detectably labelled antibody may be useful.For being in the art by the method for antibody conjugate to these agent It is known.For illustration purposes only, detectable part can be used, the labels such as radioactive atom, chromophore, fluorogen are anti- Body.The antibody of such label can be used for diagnostic techniques in vivo or in isolated test sample.They can with cell because Son, ligand, another antibody connection.It with the suitable agent for realizing antitumor action include cell factor for coupled antibody, such as Interleukin-22 (IL-2) and tumor necrosis factor (TNF)；For the photosensitizer in photodynamic therapy, including aluminum phthalocyanine tetrasulfonate (III), haematoporphyrin and phthalocyanine；Radionuclide, such as iodine -131 (131I), Yttrium-90 (90Y), bismuth -212 (212Bi), bismuth - 213 (213Bi), technetium -99m (99mTc), rhenium -186 (186Re) and rhenium-188 (188Re)；Antibiotic, such as Doxorubicin, Ah Mycin, daunorubicin, methotrexate (MTX), daunomycin, new carcinophylin and carboplatin；Bacterium, plant and other toxin, such as diphtheria poison Element, pseudomonas exotoxin A, staphylococcal enterotoxin A, abrin albumen-A toxin, ricin A (deglycosylation castor Numb toxin A and natural ricin A), TGF- alpha toxin, from Chinese cobra (naja naja atra) cytotoxin and Gelonin (a kind of phytotoxin)；Ribosome inactivating protein from plant, bacterium and fungi, such as restrictocin (one The ribosome inactivating protein kind generated by Aspergillus restrictus (Aspergillus restrictus)), saporin it is (a kind of to come from crystal soda The ribosome inactivating protein of careless (Saponaria officinalis)) and RNase；Tyrosine kinase inhibitor；ly207702 (bifluoride purine nucleosides)；Liposome (such as plasmid, the first ammonia butterfly of antisense oligonucleotides, toxin-encoding containing anti-capsule agent Purine etc.)；With other antibody or antibody fragment, such as F (ab).

Via lymphocyte antibody method (SLAM), transgenic animals and the recombinant antibodies for using hybridoma technology, selection The antibody generation that library carries out is described in greater detail below.

(1) the anti-UCH-L1 monoclonal antibody of hybridoma technology is used

Monoclonal antibody can be used multiple technologies as known in the art and (including use hybridoma, recombinant and phagocytosis Body display technology or combinations thereof) it prepares.It is, for example, possible to use hybridoma technologies to generate monoclonal antibody, the hybridoma skill Art includes as known in the art and such as those of introduction in the following: Harlow et al., Antibodies:A Laboratory Manual, the second edition, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor,1988)；Hammerling et al., In Monoclonal Antibodies and T-Cell Hybridomas, (Elsevier,N.Y.,1981).It shall yet further be noted that term " monoclonal antibody " as used herein is not limited to pass through hybridoma The antibody that technology generates.Term " monoclonal antibody " refers to the antibody from monospecific polyclonal, including any eucaryote, protokaryon life Object or phage clone, without referring to the method for generating the antibody.

It generates the method for monoclonal antibody and may include that the culture secretion present invention is anti-by the antibody that the method generates The hybridoma of body, wherein hybridoma is generated preferably through following manner: by from the animal being immunized with UCH-L1, such as greatly The splenocyte of mouse or mouse separation is merged with myeloma cell, is then screened and is secreted and can combine in the hybridoma generated by fusion The hybridoma clone of the antibody of polypeptide of the present invention.In brief, UCH-L1 antigen immune rat can be used.In preferred embodiment party In formula, UCH-L1 antigen is applied together with adjuvant to stimulate immune response.Such adjuvant includes complete or incomplete Freund Agent, RIBI (muramyl dipeptide) or ISCOM (immunostimulating complex).Such adjuvant can be by being isolated in local storage for polypeptide Protected in library polypeptide from rapid dispersion or they can secrete containing stimulation of host to macrophage and immune system Other components have the substance of the factor of chemotaxis.Preferably, if applying polypeptide, immune programme will be related to spreading In polypeptide is applied in several weeks two or more times；It is also possible, however, to use single administration polypeptide.

After with UCH-L1 antigen-immunized animal, antibody can be obtained from animal and/or generates the cell of antibody.Passing through will Animal bloodletting or execution obtain the serum containing anti-UCH-L1 antibody from animal.Blood can be used as former state by what is obtained from animal Clearly, can from serum adaptive immune immunoglobulin portion, or anti-UCH-L1 antibody can be purified from serum.In this way The serum or immunoglobulin of acquisition are polyclonal, therefore have heterogeneous series of characteristics.

Once detecting immune response, such as the antibody for being specific to antigen UCH-L1 is detected in rat blood serum, just receive Obtain rat spleen and separating Morr. cell.Then by well known technology make splenocyte and any suitable myeloma cell (such as come From the cell line SP20 that can be obtained from American Type Culture Collection (ATCC, Manassas, Va., US) Cell) fusion.Pass through the selection of limitation dilution method and clone hybridoma.Then it is measured and is hybridized by method as known in the art Secretion can be in conjunction with the cell of the antibody of UCH-L1 in tumor clone.It can be by being generated with Positive hybridoma clones immune rat Usually contain the ascites of high-level antibody.

In another embodiment, the immortal hybridoma for generating antibody can be prepared from immunity inoculation animal.Exempting from Epidemic disease inoculation after, by animal put to death and by spleen B cell as it is well known that as be fused to immortal myeloma cell.Referring to Such as Harlow and Lane, ibid.In a preferred embodiment, it is (overstepping one's bounds not secrete immunoglobulin polypeptides by myeloma cell Secreting property cell line).It is merging and after antibiotic selection, is using UCH-L1 or part thereof or express the cell screening hybridization of UCH-L1 Tumor.In a preferred embodiment, using enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA), preferably ELISA is initially screened.The example of ELISA screening is provided in PCT Publication WO 00/37504.

Selection generates the hybridoma of anti-UCH-L1 antibody, is cloned and further directed to required feature, including steady Hybridomas grew, high antibody generate and required antibody characteristic screened.Hybridoma can be immune in syngeneic animal, shortage In vivo culture and amplification in the animal (such as nude mice) of system, or in vitro culture and amplification in cell culture.Selection, gram Grand and amplified hybridization tumor method is well known to those of ordinary skill in the art.

In a preferred embodiment, hybridoma is Rat hybridoma.In another embodiment, hybridoma is non- People, non-rat species, such as mouse, sheep, pig, goat, ox or Malaysia and China generate.In another preferred embodiment again, Hybridoma is people's hybridoma, and wherein people's non-secretory myeloma is merged with the people's cell for expressing anti-UCH-L1 antibody.

The antibody fragment of identification defined epitope can be generated by known technology.For example, can be by using enzyme (such as pawpaw Protease (generate two identical Fab segments) or pepsin are (to generate F (ab')₂Segment)) to immunoglobulin molecules Proteolytic cleavage is carried out to generate Fab and F (ab') of the invention₂Segment.The F (ab') of IgG molecule₂Segment remains larger Two antigen binding sites of (" parent ") IgG molecule include light chain (containing variable light district and constant light area), heavy chain The hinge area of the formation disulfide bond of CH1 structural domain and parent's IgG molecule.Therefore, F (ab')₂Segment still is able to crosslinking antigen point Son, such as parent's IgG molecule.

(2) the anti-UCH-L1 monoclonal antibody of SLAM is used

In another aspect of the invention, using the side of the lymphocyte antibody method (SLAM) referred in the art as selection Method generates recombinant antibodies from the lymphocyte individually separated, such as U.S. Patent number 5,627,052；The PCT Publication number of opening WO 92/ 02551；With Babcook et al., described in Proc.Natl.Acad.Sci.USA, 93:7843-7848 (1996).In this method In, the unicellular of antibody interested is secreted using the screening of antigentic specificity hemolytic plaque assay method, it is immune for example originating from any The lymphocyte of animal, wherein using connector (such as biotin) by the subunit of antigen UCH-L1, UCH-L1 or its segment and silk floss Sheep red blood cell coupling, and for identifying that secretion has the unicellular of specific antibody to UCH-L1.It is interested anti-identifying After body secretory cell, heavy chain and light chain variable region cDNA are saved from cell by reverse transcriptase-PCR (RT-PCR), then this A little variable regions can in mammalian host cell, such as COS or Chinese hamster ovary celI, in constant region for immunoglobulin (example appropriate Such as human constant region) background under express.With the transfection of the immunoglobulin sequences for the amplification for being originated from the lymphocyte selected in vivo Then host cell can be further analyzed and select in vitro, such as the cell transfected by elutriation is to separate expression needle To the cell of the antibody of UCH-L1.The immunoglobulin sequences of amplification can be manipulated further in vitro, such as pass through external parent With power maturation method.See, for example, PCT Publication WO 97/29131 and PCT Publication WO 00/56772.

(3) the anti-UCH-L1 monoclonal antibody of transgenic animals is used

In yet another embodiment of the present invention, by the way that ball is immunized comprising some or all people with UCH-L1 antigen is immune The non-human animal of protein gene seat generates antibody.In one embodiment, non-human animal isTurn DNA murine, a kind of larger segment comprising human immunoglobulin gene's seat and lacks the engineered mice product that mouse antibodies generate System.See, for example, Green et al., Nature Genetics, 7:13-21 (1994) and U.S. Patent number 5,916,771,5, 939,598,5,985,615,5,998,209,6,075,181,6,091,001,6,114,598 and 6,130,364.It sees also PCT Publication WO 91/10741, WO 94/02602, WO 96/34096, WO 96/33735, WO 98/16654, WO 98/ 24893, WO 98/50433, WO 99/45031, WO 99/53049, WO 00/09560 and WO 00/37504.Transgenic mice generate fully human antibodies at proper manners people's pedigree, and generate antigentic specificity people Monoclonal antibody.Million alkali that transgenic mice passes through introducing people's heavy chain gene seat and x light chain gene seat The germline configuration YAC segment of base size and contain about 80% human antibody pedigree.Referring to Mendez et al., Nature Genetics,15:146-156(1997)；Green and Jakobovits, J.Exp.Med., 188:483-495 (1998) are public This is issued to be herein incorporated by reference.

(4) the anti-UCH-L1 monoclonal antibody in recombinant antibodies library is used

In-vitro method can also be used for preparing antibody of the invention, and wherein screening antibodies library is to identify with required UCH- The antibody of L1 binding specificity.The method of this screening about recombinant antibodies library is well known in the art, and including The method described in the following: such as U.S. Patent number 5,223,409 (Ladner et al.)；PCT Publication WO 92/18619 (Kang et al.)；PCT Publication WO 91/17271 (Dower et al.)；PCT Publication WO 92/20791 (Winter et al.)； PCT Publication WO 92/15679 (Markland et al.)；PCT Publication WO 93/01288 (Breitling et al.)；PCT is public Cloth WO 92/01047 (McCafferty et al.)；PCT Publication WO 92/09690 (Garrard et al.)；Fuchs et al., Bio/Technology,9:1369-1372(1991)；Hay et al., Hum.Antibod Hybridomas, 3:81-85 (1992)；Huse et al., Science, 246:1275-1281 (1989)；McCafferty et al., Nature, 348:552- 554(1990)；Griffiths et al., EMBO J., 12:725-734 (1993)；Hawkins et al., J.Mol.Biol., 226:889-896(1992)；Clackson et al., Nature, 352:624-628 (1991)；Gram et al., Proc.Natl.Acad.Sci.USA,89:3576-3580(1992)；Garrard et al., Bio/Technology, 9:1373- 1377(1991)；Hoogenboom et al., Nucl.Acids Res., 19:4133-4137 (1991)；Barbas et al., Proc.Natl.Acad Sci.USA,88:7978-7982(1991)；U.S. Patent Application Publication No. 2003/0186374；With PCT Publication WO 97/29131, respective content are hereby incorporated herein by.

Recombinant antibodies library may be from the subject immune with a part of UCH-L1 or UCH-L1.Optionally, recombination is anti- Body library may be from preliminary examination subject, i.e., the subject being not yet immunized with UCH-L1 is such as immune from not yet employment UCH-L1 People experimenter human antibody library.By with including people UCH-L1 peptide screening recombinant antibodies library thus to select to identify Those of UCH-L1 antibody selects antibody of the invention.Method for carrying out this screening and selection is ripe in the art Know, as described in the bibliography in previous paragraph.To select the present invention that there is particular combination affinity to UCH-L1 anti- Body, such as with specific K_offThe antibody that rate constant is dissociated from people UCH-L1 can be used surface plasma as known in the art total Method of slight selects to have required K_offThe antibody of rate constant.To select the present invention that there is specific neutralization activity to hUCH-L1 Antibody such as has specific IC₅₀Antibody, can be used as is generally known in the art for assessing standard side to the active inhibition of UCH-L1 Method.

In one aspect, the present invention relates to the separation antibodies or its antigen-binding portion thereof of a kind of combination people UCH-L1.Optionally Ground, antibody are neutralizing antibodies.In various embodiments, antibody is recombinant antibodies or monoclonal antibody.

Such as, it is possible to use various phage display methods as known in the art generate antibody.In phage display In method, functional antibodies structural domain is showed in the surface for carrying the phage particle for the polynucleotide sequence for encoding them On.This bacteriophage can be used for showing the antigen binding structure expressed from pedigree or combinatorial antibody library (such as people or muroid) Domain.It can use antigen, such as using the antigen of label or be incorporated into or be trapped in the antigen of the surface of solids or bead and select or reflect Fixed expression combines the bacteriophage of the antigen-binding domains of antigen interested.Bacteriophage used in these methods is usually filiform Bacteriophage comprising fd and M13 binding structural domain, by having recombination to be fused to phage gene III or gene VIII protein matter Fab, Fv or the stable Fv antibody domain of disulfide bond phage expression.It can be used for preparing the phage display method of antibody Example be included in method disclosed in following: Brinkmann et al., J.Immunol.Methods, 182:41-50 (1995)； Ames et al., J.Immunol.Methods, 184:177-186 (1995)；Kettleborough et al., Eur.J.Immunol, 24:952-958(1994)；Persic et al., Gene, 187:9-18 (1997)；Burton et al., Advances in Immunology,57:191-280(1994)；PCT Publication WO 92/01047；PCT Publication WO 90/02809；WO 91/ 10737；WO 92/01047；WO 92/18619；WO 93/11236；WO 95/15982；WO 95/20401；And United States Patent (USP) Numbers 5,698,426,5,223,409,5,403,484,5,580,717,5,427,908,5,750,753,5,821,047,5, 571,698,5,427,908,5,516,637,5,780,225,5,658,727,5,733,743 and 5,969,108.

It, can be from bacteriophage separation antibody code area and for producing after phage selection as described in above-mentioned bibliography Raw complete antibody, including human antibody or any other required antigen-binding fragment, and expressed in any required host, The host includes mammalian cell, insect cell, plant cell, yeast and bacterium, such as described below.For example, Method as known in the art can be used to use to be used to recombinate and generate Fab, Fab' and F (ab')₂The technology of segment, such as with Technology disclosed in lower: PCT Publication WO 92/22324；Mullinax et al., BioTechniques, 12 (6): 864- 869(1992)；Sawai et al., Am.J.Reprod.Immunol, 34:26-34 (1995)；With Better et al., Science, 240:1041-1043(1988).The example that can be used for generating the technology of scFv and antibody includes U.S. Patent number 4,946,778 With 5,258,498；Huston et al., Methods in Enzymology, 203:46-88 (1991)；Shu et al., Proc.Natl.Acad,Sci.USA,90:7995-7999(1993)；With Skerra et al., Science, 240:1038-1041 (1988) technology described in.

It is big for screening as is generally known in the art as the alternative solution for screening recombinant antibodies library by phage display Other methods of type combinatorial libraries can be applied to identify antibody of the present invention.As PCT Publication WO 98/31700 (Szostak and Roberts) and Roberts and Szostak, Proc.Natl.Acad.Sci.USA, described in 94:12297-12302 (1997), A kind of alternative expression system is the expression system that recombinant antibodies library is expressed as RNA- protein fusions.In this system In, by being translated in vitro in a kind of synthesis mRNA of the end 3' with puromycin (peptidyl receptor antibiotic) come in mRNA Covalent fusions are generated between the peptide or protein matter encoded with it.It therefore, can be based on encoded peptide or protein matter (such as antibody Or part thereof) characteristic (such as combination of antibody or part thereof and dual specificity antigen), from the complex mixture (example of mRNA Such as combinatorial libraries) enrichment specific mrna.Recombination form as described above (such as in mammalian host cell) can be passed through The nucleic acid sequence of encoding antibody recycled from the such library of screening or part thereof is expressed, and in addition can be by having drawn to mutation Enter to the mRNA- peptide fusions in the sequence initially selected and carries out more wheels screening or by as described above for recombinant antibodies Other methods of external affinity maturation be subjected to further affinity maturation.The preferred embodiment of this method is PROfusion Display technique.

In another way, antibody can also be used yeast display method as known in the art to generate.In yeast exhibition Show in method, genetic method is used for antibody domain drift bolt to yeast cell wall and them is made to be showed in yeast surface On.Specifically, this primary yeast can be used for showing the antigen knot expressed from pedigree or combinatorial antibody library (such as people or muroid) Close structural domain.The example that can be used for preparing the yeast display method of antibody includes the United States Patent (USP) being hereby incorporated herein by Method disclosed in number 6,699,658 (Wittrup et al.).

D, the generation of UCH-L1 antibody is recombinated

Antibody can be generated by any of many technologies as known in the art.For example, from host cell expression, In the expression vector of encoding heavy chain and light chain is transfected into host cell by standard technique.Various forms of terms " transfection " It is intended to cover commonly used in the extensive multiple technologies that introduce exogenous DNA in protokaryon or eukaryotic host cell, such as electroporation, Calcium phosphate precipitation, the transfection of DEAE- polydextrose etc..While it may be possible to it is anti-to express the present invention in protokaryon or eukaryotic host cell Body, but antibody preferably is expressed in eukaryocyte and most preferably mammalian host cell, because such eukaryocyte (and especially It is mammalian cell) correct folding and resisting with immunologic competence are more likely assembled and secreted compared to prokaryotic cell Body.

Exemplary mammals host cell for expressing recombinant antibodies of the invention includes Chinese hamster ovary cell (Chinese hamster ovary celI) (including Urlaub and Chasin, Proc.Natl.Acad.Sci.USA, described in 77:4216-4220 (1980) Dhfr-CHO cell, be used together with DHFR selectable marker, such as such as Kaufman and Sharp, J.Mol.Biol., Described in 159:601-621 (1982)；NS0 myeloma cell；COS cell and SP2 cell.When by the recombination of encoding antibody genes When expression vector is introduced into mammalian host cell, allow antibody thin in host by the way that one section of host cell culture to be enough Time in born of the same parents in expression or the culture medium for more preferably growing antibody-secreting to host cell generates antibody.Antibody can To use Standard protein purification method to recycle from culture medium.

Host cell can also be used for generating functional antibody fragment, such as Fab segment or scFv molecule.It should be appreciated that can To carry out modification to above procedure.For example, it may be possible to it is desirable that with the light chain of coding antibody of the present invention and/or the function of heavy chain The DNA transfection host cell of energy property segment.Recombinant DNA technology can also be used for removing some or all codings and non-binding interested The DNA of any of light chain and heavy chain or both necessary to antigen.Antibody of the present invention, which is also covered by, truncates DNA molecular by this class The molecule of expression.In addition, double function can be generated by being crosslinked antibody of the present invention and secondary antibody with standard chemical cross-linking method Energy antibody, one of heavy chain and a light chain are antibody of the present invention (combining people UCH-L1) and another heavy chain and another light chain There is specificity to the antigen of inhuman UCH-L1.

In an optimum decision system for recombinantly expressing antibody of the present invention or its antigen-binding portion thereof, pass through calcium phosphate The recombinant expression carrier of both encoding antibody heavy and antibody light chain is introduced into dhfr-CHO cell by the infection protocol of mediation.In In recombinant expression carrier, heavy chain of antibody and light chain gene is made to be each operably linked to cmv enhancer/AdMLP promoter tune Control element is to drive gene height to transcribe.Recombinant expression carrier allows with DHFR gene using methotrexate (MTX) selection/expansion Increase to select the Chinese hamster ovary celI transfected with carrier.Selected transformant host cell is cultivated to allow to express heavy chain of antibody Complete antibody is recycled with light chain and from culture medium.Standard molecular biological technique is used to prepare recombinant expression carrier, is turned Host cell, selection transformant, culture host cell are contaminated, and recycles antibody from culture medium.Further, the present invention mentions The method for the recombinant antibodies for having supplied synthesis of the invention, the method are thin by cultivating host of the invention in being suitble to culture medium Born of the same parents carry out until synthesizing recombinant antibodies of the invention.The method may further include separates recombinant antibodies from culture medium.

(1) humanized antibody

" humanized antibody " can be antibody or its variant, derivative, analog or part, immunospecifically combine Antigen interested and include the substantially amino acid sequence with human antibody the frame area (FR) and substantially have non-human antibody Amino acid sequence complementary determining region (CDR).Humanized antibody can come from non-human species in conjunction with required antigen, have Complementary determining region (CDR) of the one or more from non-human species and the framework region from human immunoglobulin molecule.

As used herein, the term under the background of CDR " substantially " refers to the ammonia of amino acid sequence Yu non-human antibody CDR Base acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical CDR.Humanized antibody includes substantially complete At least one and usual two variable domains (Fab, Fab', the F (ab') in portion₂, FabC, Fv), wherein completely or generally complete Portion's CDR region corresponds to those of non-human immunoglobulin (i.e. donor antibody) CDR region and completely or generally whole framework regions It is those of human immunoglobulin(HIg) consensus sequence.According on one side, humanized antibody also includes constant region for immunoglobulin (Fc) at least part of (constant region of usual human immunoglobulin(HIg)).In some embodiments, humanized antibody contains gently Both variable domains of chain and at least heavy chain.Antibody also may include the CH1, hinge, the area CH2, CH3 and CH4 of heavy chain.In In some embodiments, humanized antibody has contained only humanization light chain.In some embodiments, humanized antibody contains only someone Source heavy chain.In certain embodiments, humanized antibody only humanization variable domains containing light chain and/or heavy chain.

Humanized antibody can be selected from any classification of immunoglobulin, including IgM, IgG, IgD, IgA and IgE, and Any isotype, including but not limited to IgG1, IgG2, IgG3 and IgG4.Humanized antibody may include from more than one class Other or isotype sequence, and it is required to optimize that the specific constant domain of the choice of technology well known in the art can be used Effector function.

The framework region and CDR region of humanized antibody are not necessarily to correspond precisely to parental array, such as can be by replacing, being inserted into Or/or lack at least one amino acid residue and to carry out mutagenesis to donor antibody CDR or shared frame the CDR so that at the site Or Framework residues do not correspond to donor antibody or shared frame.However, in one embodiment, such mutation will not be extensive 's.In general, at least 90%, at least 95%, at least 98% or at least 99% humanized antibody residue will correspond to parent FR and Those of CDR sequence.As used herein, term " shared frame " refers to the framework region in shared immunoglobulin sequences.Such as this Used in text, term " shared immunoglobulin sequences " refers to by amino acid the most common in associated immunoglobulin sequence family The sequence that (or nucleotide) is formed is (see, for example, Winnaker, From Genes to Clones (Verlagsgesellschaft,Weinheim,Germany 1987)).In immunoglobulin class, in consensus sequence Each position is occupied by the amino acid for most often appearing in that position in family.If two amino acid continually occur on an equal basis, It so may include any one in consensus sequence.

Humanized antibody can be designed so that the undesired immune response to rodent anti-human antibody minimizes, this is limited Duration and the validity for the treatment of use of those parts in human recipient are made.Humanized antibody can have from non- People source introduces one or more of amino acid residues.These non-human residues commonly referred to as " input " residue, usually It is derived from variable domains.It can be by replacing the corresponding sequence of human antibody to carry out humanization with hypervariable region sequence.Therefore, such " humanization " antibody is chimeric antibody, wherein significantly less than complete people's variable domains by the corresponding sequence from non-human species Column replace.For example, with reference to U.S. Patent number 4,816,567, disclosure is hereby incorporated herein by.Humanization Antibody can be human antibody, and some of some hypervariable region residues and possible some FR residues are by the class in rodent animal antibody Replace like the residue in site.Any known method can be used and carry out humanization or engineering to antibody of the present invention, such as but Be not limited to U.S. Patent number 5,723,323,5,976,862,5,824,514,5,817,483,5,814,476,5,763,192, 5,723,323、5,766,886、5,714,352、6,204,023、6,180,370、5,693,762、5,530,101、5,585, 089, method described in 5,225,539 and 4,816,567.

Humanized antibody can retain high-affinity and other advantageous biological natures to UCH-L1.Parent can be used The threedimensional model of sequence and humanized sequence is prepared by the analysis method of parental array and various conceptual humanized products Humanized antibody.Three dimensional immunoglobulin model is usually obtainable.Illustrate and show selected candidate immunoglobulin sequences sequence The computer program of possibility three-dimensional conformation structure be obtainable.It is immune in candidate that these inspections shown allow to analyze residue The residue that possibility effect in the function of globin sequence, i.e. analyzing influence candidate immunoglobulin sequences combine the ability of its antigen. In this way, FR residue can be selected and merged from recipient and list entries, thus the antibody characteristic needed for realizing, such as The increased affinity to UCH-L1.In general, some hypervariable region residues directly and most may substantially be related to influence antigen knot It closes.

As the alternative solution of humanization, it can produce human antibody (referred to herein as " fully human antibodies ").For example, Can via PROfusion and/or yeast the relevant technologies the isolating human antibodies from library.Transgenic animals (example can also be generated Such as, mouse), the transgenic animals can be generating people there is no endogenous immunoglobulin generates after immune The full pedigree of antibody.For example, antibody heavy chain joining region (the J in chimeric and germ line mutant mice_H) homozygous deletion of gene leads Complete inhibition endogenous antibody is caused to generate.Transfer of human germline immunoglobulin's Gene Array in this germ line mutant mice will Cause to generate human antibody after antigen stimulation.Humanization or fully human antibodies can according to U.S. Patent number 5,770,429,5, 833,985、5,837,243、5,922,845、6,017,517、6,096,311、6,111,166、6,270,765、6,303, 755,6,365,116,6,410,690,6,682,928 and 6, prepared by method described in 984,720, the patent is respective Content is hereby incorporated herein by.

E, anti-UCH-L1 antibody

Above-mentioned technology can be used and generate anti-UCH-L1 antibody using routine techniques as known in the art.Some In embodiment, anti-UCH-L1 antibody can be unconjugated UCH-L1 antibody, such as can be from the following UCH-L1 antibody obtained: United State Biological (catalog number (Cat.No.): 031320), Cell Signaling Technology (catalog number (Cat.No.): 3524), Sigma-Aldrich (catalog number (Cat.No.): HPA005993), Santa Cruz Biotechnology, Inc. (catalog number (Cat.No.): Sc-58593 or sc-58594), R&D Systems (catalog number (Cat.No.): MAB6007), Novus Biologicals (catalog number (Cat.No.): NB600-1160), Biorbyt (catalog number (Cat.No.): orb33715), Enzo Life Sciences, Inc. (catalog number (Cat.No.): ADI-905- 520-1), Bio-Rad (catalog number (Cat.No.): VMA00004), BioVision (catalog number (Cat.No.): 6130-50), Abeam (catalog number (Cat.No.): Ab75275 or ab104938), Invitrogen Antibodies (catalog number (Cat.No.) 480012), ThermoFisher Scientific (catalog number (Cat.No.): MA1-46079, MA5-17235, MA1-90008 or MA 1-83428), EMD Millipore (mesh MABN48) or Sino Biological Inc. (catalog number (Cat.No.): 50690-R011) record number:.Anti- UCH-L1 antibody can be with fluorescence Group's conjugation, such as can be from BioVision (catalog number (Cat.No.): 6960-25) or Aviva Systems Biology (catalog number (Cat.No.) OAAF01904-FITC) the UCH-L1 antibody of the conjugation obtained.

11, the modification of method

The presence of analytes of interest analytes (UCH-L1) present in the disclosed product of random sample really or the method for amount can be such as these Described in text.The method can also be adjusted according to other methods for analysis of analytes.The example packet of well known modification Include but be not limited to immunoassay, such as sandwich immunoassays (such as monoclonal-monoclonal sandwich immunoassays, Dan Ke Grand-polyclonal sandwich immunoassays, including enzyme detection (enzyme immunoassay (EIA) or enzyme-linked immunosorbent assay) (ELISA), Reverse transcriptase immunoassay (such as forward and reverse), enzyme expand immunoassay (EMIT), competitive Binding assay, bioluminescence resonance energy transfer (BRET), a step antibody test measuring method, homogeneous measuring method, heterogeneous measurement Method captures measuring method etc. immediately.

A, immunoassay

Can in immunoassay using UCH-L1 antibody analysis analytes of interest analytes and/or its peptide or segment (such as UCH-L1, and/or its peptide or its segment, i.e. UCH-L1 segment).Antibody can be used and detect and analyte (such as UCH- L1 specific binding) determines the presence or amount of analyte (such as UCH-L1).For example, antibody or its antibody fragment can be special Anisotropic binding analysis object (such as UCH-L1).If desired, one or more antibody commercially available can be obtained with one or more Monoclonal/the polyclonal antibody obtained is applied in combination.Such antibody can from such as R&D Systems, Inc. (Minneapolis, MN it) is obtained with the company of Enzo Life Sciences International, Inc. (Plymouth Meeting, PA).

Immunoassay can be used easily in the presence of analyte present in vivo sample (such as UCH-L1) or amount Determination, such as sandwich immunoassays (such as monoclonal-monoclonal sandwich immunoassays, the polyclonal sandwich immunoassay of monoclonal- Measuring method, including radioactive isotope detection (radioimmunoassay (RIA)) and enzyme detect (enzyme immunoassay (EIA) or Enzyme-linked immunosorbent assay (ELISA) (such as Quantikine ELISA assays, R&D Systems, Minneapolis,MN)).The example for the point-of-care device that can be used is i-(Abbott,Laboratories, Abbott Park,IL).The other methods that can be used for example including chemiluminescence particle immunoassay, especially withThe method of automatic analyzer (Abbott Laboratories, Abbott Park, IL).Other methods It is (such as anti-using analysis resistant object including such as mass spectrography and immunohistochemical method (such as slice for self-organizing biopsy) UCH-L1) antibody (monoclonal, polyclonal, chimeric, humanization, people etc.) or its be directed to analyte (such as UCH-L1) antibody Segment.Other detection methods include such as U.S. Patent number 6,143,576,6,113,855,6,019,944,5,985,579,5, 947,124、5,939,272、5,922,615、5,885,527、5,851,776、5,824,799、5,679,526、5,525,524 With 5, described in 480,792 those, the patent is respectively incorporated herein in its entirety by reference.Antibody and analyte (example Such as UCH-L1) specific immunity combine can be via the direct label for being attached to antibody, such as fluorescence or Luminous label, metal And radionuclide, or via indirect labelling, such as alkaline phosphatase or horseradish peroxidase are detected.

The use of immobilized antibody or its antibody fragment can be incorporated into immunoassay.Antibody immobilization can be existed On a variety of supports, such as magnetic or chromatography matrix particle, the surface of assay plate (such as microtiter well), solid matrix materials Piece etc..Measurement strip can be prepared by the way that antibody or Multiple Antibodies to be coated on solid support with array manner.Then may be used It is such as aobvious this item to be immersed in test sample and is handled rapidly by washing and detecting step to generate measurable signal The point of color.

Homogeneous form can be used.For example, preparing mixture after obtaining test sample from subject.Mixture, which contains, to be wanted Test sample, the first specific binding partner and the second specific binding partner of analysis and assessment object (such as UCH-L1). Test sample, the first specific binding partner and the second specific binding partner are added to form the sequence of mixture not It is crucial.Contact test sample with the first specific binding partner and the second specific binding partner simultaneously.In some realities It applies in mode, any UCH-L1 contained in the first specific binding partner and test sample can form the first specificity knot It closes gametophyte-analyte (such as UCH-L1)-antibody complex and the second specific binding partner can form the first spy The-the second specific binding partner compound of specific binding partner-analytes of interest analytes (such as UCH-L1).In some implementations In mode, any UCH-L1 contained in the second specific binding partner and test sample can form the second specific binding It is special that gametophyte-analyte (such as UCH-L1)-antibody complex and the first specific binding partner can form first Property binding partners-analytes of interest analytes (such as UCH-L1)-the second specific binding partner compound.First specificity knot Closing gametophyte can be analysis resistant object antibody (for example, combining has the continuous amino at least three (3) comprising SEQ ID NO:1 The anti-UCH-L1 antibody of the epitope of the amino acid sequence of acid).Second specific binding partner can be analysis resistant object antibody (example There is the anti-UCH-L1 of the epitope of amino acid sequence of at least three (3) continuous amino acids comprising SEQ ID NO:1 as combined Antibody).In addition, the second specific binding partner marks with detectable label as described above or contains the detectable mark Note.

Heterogeneous form can be used.For example, preparing the first mixture after obtaining test sample from subject.It is described Mixture contains the test sample and the first specific binding partner for wanting analysis and assessment object (such as UCH-L1), wherein first is special Any UCH-L1 contained in specific binding partner and test sample forms the first specific binding partner-analyte (example Such as UCH-L1)-antigenic compound.First specific binding partner can be analysis resistant object antibody (such as combine have comprising The anti-UCH-L1 antibody of the epitope of the amino acid sequence of at least three (3) continuous amino acids of SEQ ID NO:1).Addition test Sample and the first specific binding partner are not critical with the sequence for forming mixture.

First specific binding partner can be immobilized in solid phase.For immunoassay (for the first specificity Binding partners and the second optional specific binding partner) in solid phase can be any solid phase being known in the art, Such as, but not limited to magnetic-particle, bead, test tube, microtiter plate, colorimetric cylinder, film, scaffold molecule, film, filter paper, disc and Chip.It is in those of bead embodiment in solid phase, bead can be magnetic beads or magnetic-particle.Magnetic beads/particle It can be ferromagnetic, ferrimagnetism, paramagnetic, superparamagnetism or ferrofluid.Exemplary ferromagnetic material includes Fe、Co、Ni、Gd、Dy、CrO₂, MnAs, MnBi, EuO and NiO/Fe.The example of ferrimagnetic material includes NiFe₂O₄、 CoFe₂O₄、Fe₃O₄(or FeO'Fe₂O₃).Bead can have magnetic solid core part and by one or more non magnetic Layer surrounds.Optionally, magnetic part can be the layer around non-magnetic core.Immobilization thereon has the first specific binding members Solid support can be in a dry form or with fluid storage.Magnetic bead is having the first specific binding members with immobilization thereon Magnetic bead contact before or after can be subjected to magnetic field.

Formed containing the first specific binding partner-analyte (such as UCH-L1) antigenic compound mixture it Afterwards, any unbonded analyte (such as UCH-L1) is removed from compound using any technology being known in the art.Example Such as, unbonded analyte (such as UCH-L1) can be removed by washing.However, it is desirable to the first specific binding is matched Even body more than the amount of any analyte (such as UCH-L1) in the presence of test sample to exist, so that being present in test specimens All analytes (such as UCH-L1) in product are all combined by the first specific binding partner.

After removing any unbonded analyte (such as UCH-L1), the second specific binding partner is added to Matched in mixture with forming the specific binding of the first specific binding partner-analytes of interest analytes (such as UCH-L1)-the second Even nanocrystal composition.Second specific binding partner can be analysis resistant object antibody (such as combine have comprising SEQ ID NO:1 At least three (3) continuous amino acids amino acid sequence epitope anti-UCH-L1 antibody).In addition, the second specific binding Gametophyte marks with detectable label as described above or contains the detectable label.

The use of immobilized antibody or its antibody fragment can be incorporated into immunoassay.Antibody immobilization can be existed On a variety of supports, the modified latex of such as magnetism or chromatography matrix particle (such as magnetic beads), latex particle or surface Grain, polymer or thin polymer film, plastics or plastic film, planar substrate, assay plate (such as microtiter well) surface, solid Body host material piece etc..Measurement can be prepared by the way that antibody or Multiple Antibodies to be coated on solid support with array manner Item.Then this item can be immersed in test sample and is handled rapidly by washing and detecting step to generate and can measure letter Number, the point such as to develop the color.

(1) sandwich immunoassays

Sandwich immunoassays measure two layers of antibody (i.e. at least one capture antibody) and (the i.e. at least one inspection of detection antibody Survey antibody) between amount of antigen.It captures on antibody and detection antibodies bind antigen, such as analytes of interest analytes (such as UCH-L1) Different epitopes.It is desirable that capture antibody and epitope combination will not Interference Detection antibody and epitope combination.Monoclonal is more Clonal antibody can be employed as capture antibody and detection antibody in sandwich immunoassays.

In general, being separated using at least two antibody and the analyte (such as UCH-L1) in quantitative test sample.More Body, certain epitopes at least two antibody binding assay objects (such as UCH-L1) are claimed to form immune complex For " sandwich ".(these antibody are normal for the analyte (such as UCH-L1) that can be used to capture by one or more antibody in test sample Referred to as one or more " capture " antibody) and one or more antibody can be used to make can detect and (can quantify) label combination Sandwich (these antibody are frequently referred to one or more " detection antibody ").In sandwich assay, the combination of antibody and its epitope is managed Think ground will not because measurement in any other corresponding epitope of antibody in conjunction with due to weaken.Select antibody so that with it is doubtful containing One or more first antibodies of the test sample contact of analyte (such as UCH-L1) are not identified with second or subsequent antibody The all or part of of epitope combine, to interfere one or more second detections antibody binding assay object (such as UCH-L1) Ability.

Antibody can be used as first antibody in the immunoassay.Table on antibody mediated immunity specific binding assay object Position (such as UCH-L1).Other than antibody of the invention, the immunoassay may include secondary antibody, be immunized special Property combine by first antibody identify or combine epitope.

The doubtful test sample containing analyte (such as UCH-L1) can simultaneously or sequentially be caught at least one first It obtains antibody (or a variety of first capture antibody) and at least one second detects antibody contact.In sandwich assay formats, exist first Allow to be formed under conditions of first antibody-analyte (such as UCH-L1) antigenic compound make it is doubtful containing analyte (such as UCH-L1 test sample) is contacted with the first capture antibody of at least one specific binding defined epitope.If using being more than A kind of capture antibody then forms more than first kind of capture antibody-UCH-L1 antigenic compound.In sandwich assay, antibody, preferably At least one capture antibody, with the maximum amount of mole of mistake relative to analyte (for example, UCH-L1) expected in test sample Amount amount uses.For example, every ml particle coating buffer can be used about 5 μ g/ml to about 1mg/ml antibody.

I, anti-UCH-L1 captures antibody

Optionally, before contacting test sample at least one the first capture antibody, at least one capture antibody can It is bound to solid support, the solid support is conducive to separate first antibody-analyte (such as UCH- from test sample L1) compound.Any solid support as known in the art can be used, be including but not limited to made of polymer material, In the solid support in hole, pipe or bead (such as particle) form.One (or more) kinds of antibody can by absorption, by using The covalent bonding of chemical coupling agent or by other means as known in the art in conjunction with solid support, precondition is this Kind combination will not interfere the ability of antibody binding assay object (such as UCH-L1).In addition, if need, it can be by solid support Derivatization is to allow and the various functional group reactions on antibody.This derivative words are needed using certain coupling agents, such as but unlimited In maleic anhydride, n-hydroxysuccinimide and 1- ethyl -3- (3- dimethylaminopropyl) carbodiimides.

Hereafter the test sample by doubtful containing analyte (such as UCH-L1) is incubated for allow to be formed the first capture antibody (or a variety of capture antibody)-analyte (such as UCH-L1) compound.Incubation can be at the pH of about 4.5 to about 10.0, at about 2 DEG C To at a temperature of about 45 DEG C, and continue at least about (1) minute to about 18 (18) hours, about 2-6 minutes, about 7-12 points Clock, about 5-15 minutes or about 3-4 minutes period carry out.

Ii, detection antibody

After forming first/a variety of capture antibody-analyte (such as UCH-L1) compounds, then make compound and at least One kind second, which detects antibody contact, (is allowing to be formed first/Multiple Antibodies-analyte (such as UCH-L1) antigen-secondary antibody Under conditions of compound).In some embodiments, test sample is contacted with detection antibody simultaneously with capture antibody.If First antibody-analyte (such as UCH-L1) compound is contacted with more than one detection antibody, then formation first/a variety of captures Antibody-analyte (such as UCH-L1)-Multiple Antibodies detect compound.As first antibody, when make at least second (and after It is continuous) antibody and when first antibody-analyte (such as UCH-L1) complex contacts, it is incubated under conditions of similar with the above It educates and is formed needed for first/Multiple Antibodies-analyte (such as UCH-L1)-the second/Multiple Antibodies compound for a period of time.It is excellent Selection of land, at least one secondary antibody contain detectable label.Forming first/Multiple Antibodies-analyte (such as UCH-L1)-the Prior to, concurrently with, or after two/Multiple Antibodies compound, detectable label can be in conjunction at least one secondary antibody.It can be used Any detectable label as known in the art.

Chemiluminescence assay can be according to Adamczyk et al., (2006) Anal.Chim.Acta 579 (1): 61-67 Described in method carry out.Although any suitable measuring method form can be used, microplate chemiluminescence meter (Mithras LB- 940, Berthold Technologies U.S.A., LLC, Oak Ridge, TN) make it possible to quickly measure multiple small sizes Sample.When using 96 hole black polystyrene microwell plate (Costar#3792), chemiluminescence meter can be infused equipped with multiple reagents Emitter.Each sample can be added in individual hole, then simultaneously/successively addition is as true by used type institute Fixed other reagents.It is desirable that avoiding being formed using the neutrality or the pseudobase in alkaline solution of acridine aryl ester, such as pass through acid Change.Then chemiluminescence response is hole-specifically recorded.In this regard, it depends on adding with recording the time portion of chemiluminescence response Delay between reagent adding and used specific acridine.

Add test sample and specific binding partner) to form the sequence of the mixture for chemiluminescence assay It is not critical.If the first specific binding partner is detectably marked with acridine compounds, is formed and detectably marked The first specific binding partner-UCH-L1 antigenic compound.Optionally, if simultaneously using the second specific binding partner And second specific binding partner detectably marked with acridine compounds, then formed detectably mark first specificity The-the second specific binding partner compound of binding partners-analyte (such as UCH-L1).Any unbonded specificity knot Close that gametophyte (no matter marking or unlabelled) all can be used any technology as known in the art (such as washing) from mixture Middle removing.

Hydrogen peroxide can generate in situ in the mixture, or before adding above-mentioned acridine compounds, simultaneously or it There is provided or be supplied to afterwards mixture.Hydrogen peroxide can (such as will be in many ways apparent to those skilled in the art Mode) generation in situ.

It is alternatively possible to which hydrogen peroxide source is simply added to mixture.For example, hydrogen peroxide source can be known contain There are the one or more buffers or other solution of hydrogen peroxide.In this regard, can simply to add hydrogen peroxide molten Liquid.

While adding at least one alkaline solution into sample or later, generates instruction the existing of UCH-L1 and examine Survey signal, i.e. chemiluminescence signal.Alkaline solution contains at least one alkali and has and is greater than or equal to 10, is preferably greater than or waits In 12 pH.The example of alkaline solution includes but is not limited to sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonium hydroxide, hydroxide Magnesium, sodium carbonate, sodium bicarbonate, calcium hydroxide, calcium carbonate and calcium bicarbonate.The amount for the alkaline solution being added in sample depends on The concentration of alkaline solution.Based on the concentration of alkaline solution used, those skilled in the art, which can easily determine, to be added in sample Alkaline solution amount.It can be using other labels in addition to chemiluminescent labeling.For example, can be marked using enzyme (including But it is not limited to alkaline phosphatase).

The chemiluminescence signal or other signals that routine techniques detection well known by persons skilled in the art generates can be used. Based on the intensity for generating signal, the amount of the analytes of interest analytes (such as UCH-L1) in sample can be quantified.Specifically, in sample The amount of analyte (such as UCH-L1) is proportional to the intensity of signal is generated.The amount of existing analyte (such as UCH-L1) can lead to It crosses the amount of produced light compared with the standard curve of analyte (such as UCH-L1) or by being quantified compared with reference standard. The known dense of analyte (such as UCH-L1) can be used by mass spectrum, gravimetric method and other technologies as known in the art The serial dilution or solution of degree generates standard curve.

(2) positive Reverse transcriptase measuring method

In positive competition model, using the analytes of interest analytes of the label of known concentration (such as with fluorescent marker (one The label kind being attached with cracking joint) etc. analyte (such as UCH-L1)) aliquot and test sample in it is interested Analyte (such as UCH-L1) competitive binding analytes of interest analytes antibody (such as UCH-L1 antibody).

In positive competition assay, the specific binding partner (such as antibody) of immobilization can sequentially or simultaneously with survey The analytes of interest analytes of test agent and label, its analytes of interest analytes segment or the contact of analytes of interest analytes variant.Interested point Analysis object peptide, analytes of interest analytes segment or analytes of interest analytes variant can be marked with any detectable label, including by with can Crack the detectable label of the label composition of connector attachment.It, can be by antibody immobilization on solid support in the measurement. It is alternatively possible to by antibody and the antibody being immobilized on solid support (such as particle or planar substrate), such as anti-species Antibody coupling.

By the analytes of interest analytes, test sample and antibody of label those of described in the sandwich assay formats above in conjunction It is incubated under conditions of similar.Then it can produce two different kinds of antibody-analytes of interest analytes compound.Specifically, it produces One of raw antibody-analytes of interest analytes compound contains detectable label (such as fluorescent marker etc.), and another antibody-sense Interest analysis object is free of detectable label.Antibody-analytes of interest analytes compound can with but need not quantitative detectable label it It is preceding to be separated with the rest part of test sample.No matter antibody-analytes of interest analytes compound whether its remaining part with test sample Point separation, all and then quantitatively in antibody-analytes of interest analytes compound detectable marker amount.Then test specimens can be determined Analytes of interest analytes (the relevant analytes of interest analytes of such as film, soluble analytes of interest analytes, soluble analysis interested in product The segment of object, variant (film relevant or soluble analytes of interest analytes) of analytes of interest analytes or any combination thereof) concentration, Such as it is as described above.

(3) reversed Reverse transcriptase measuring method

In reversed competition assay, the analytes of interest analytes (such as UCH-L1) of immobilization can sequentially or simultaneously with survey Test agent and at least one labelled antibody contact.

Analytes of interest analytes can be with solid support, such as solid support above in association with sandwich assay formats discussion In conjunction with.

By the antibody of the analytes of interest analytes of immobilization, test sample and at least one label in sandwich survey above in conjunction It is incubated for similar under conditions of those of described in setting formula.Then it is compound to generate two different kinds of analytes of interest analytes-antibody Object.Specifically, one of analytes of interest analytes-antibody complex of generation is immobilized and (such as glimmering containing detectable label Signal etc.), and another analytes of interest analytes-antibody is not immobilized and without detectable label.By this field The technology known, is such as washed, and the sense that unlockedization is removed from analytes of interest analytes-antibody complex presence of immobilization is emerging The rest part of interesting analyte-antibody complex and test sample.Once the analytes of interest analytes antibody for removing unlockedization is multiple Close the amount that object then quantifies detectable label in analytes of interest analytes-antibody complex analyte of immobilization after cracking label. The quantity of comparison detectable label as described above be may then pass through to determine the concentration of analytes of interest analytes in test sample.

(4) one step immunoassays or " capture immediately " measuring method

In capture immunoassay immediately, solid matrix is pre-coated with fixative.By capturing agent, analyte (example Such as UCH-L1) and detection agent be added to solid matrix together, be washing step later, then detect.Capturing agent, which can combine, to be divided It analyses object (such as UCH-L1) and includes the ligand for fixative.Capturing agent and detection agent can be antibody or as described herein Or any other part as known in the art that can be captured or detect.Ligand can wrap peptide tag and fixative can wrap Containing anti-peptide tag antibody.Optionally, ligand and fixative, which can be, to be combined together, for use in instant capture measuring method Any reagent to (such as specific binding pair, and as known in the art other reagents to).It can measure more than one The analyte of kind.In some embodiments, antigen applying solid matrix can be used, and analyte to be analyzed is antibody.

In certain other embodiments, in a step immunoassay or " capture immediately ", uses and be pre-coated with The solid support (such as particle) of fixative (biotin, streptavidin etc.) and at least first specificity knot Synthesis person and the second specific binding members (being used separately as capture reagent and detection reagent).First specific binding members include For fixative ligand (for example, if the fixative on solid support is streptavidin, then the first specificity Ligand on binding members can be biotin) and herein in connection with analytes of interest analytes (such as UCH-L1).Second specificity knot Synthesis person includes detectable label and combines analytes of interest analytes (such as UCH-L1).It can be different by solid support and first Property binding members and the second specific binding members (sequentially or simultaneously) are added in test sample.First specific binding members On ligand in conjunction with the fixative on solid support, formed solid support/first specific binding members compound.It deposits It is that any analytes of interest analytes in sample is solid to be formed in conjunction with solid support/first specific binding members compound Body support/the first specific binding members/analyte complex.Second specific binding members and solid support/the first Specific binding members/analyte complex combines, and detects detectable label.It can use before testing optionally Washing step.In some embodiments, in a step measuring method, more than one analyte can be measured.Certain other It, can be using the specific binding members more than two kinds in embodiment.In certain other embodiments, it can add a variety of Detectable label.It in certain other embodiments, can detecte a variety of analytes of interest analytes, or measure, determine or comment Estimate their amount, level or concentration.

Immediately the use of capture measuring method can be carried out with diversified forms as described herein, and be in the art Know.It for example, the form can be sandwich assay as described above, but optionally can be competition assay, can adopt With single specificity binding members or use all other modifications as is known.

12, other factors

The method of as described above diagnosis, prognosis and/or assessment may further include diagnosed using other factors, Prognosis and assessment.In some embodiments, Glasgow final result scale of Golmud-Lhasa Pipeline or extension can be used (GOSE) carry out Diagnosis of Trauma cerebral injury.Also it can be used alone or other tests, amount be applied in combination with Golmud-Lhasa Pipeline Table or index.One example is auspicious autumn Loews A meter Ge Si scale (Ranchos Los Amigos Scale).Auspicious Qiu Luosia Meter Ge Si scale measurement consciousness, cognition, behavior and the level interacted with environment.Auspicious autumn Loews A meter Ge Si scale includes: I Grade: reactionless；II grades of systemic reactions；III level: local reaction；IV grades: chaotic-restless；V grades: chaotic-inappropriate；VI grades: It is chaotic-appropriate；VII grades: automatic-appropriate；With VIII grades: purposeful-appropriate.

13, sample

In some embodiments, in human experimenter by being shaken by body, closed or open head is caused to be created Passivity impact, one or many tumbles, explosion or the shock wave or other types of that the external mechanical force of wound or other power generate Sample is obtained later to the damage on head caused by blunt forces wound.In some embodiments, in human experimenter's intake or cruelly The combination for being exposed to chemical substance, toxin or chemical substance and toxin obtains sample later.The reality of such chemical substance and/or toxin Example includes fire, mould, asbestos, insecticide and insecticide, organic solvent, paint, glue, gas (such as carbon monoxide, vulcanization Hydrogen and cyanide), organic metal (such as methyl mercury, lead tetraethide and organotin) and/or one or more drug abuse.One In a little embodiments, sample is from suffering from autoimmune disease, metabolic disorder, brain tumor, anoxic, one or more viruses, meninx The human experimenter of scorching, hydrocephalus or combinations thereof obtains.

In another embodiment again, the sample that method described herein uses be can be also used for by using hereafter institute The anti-UCH-L1 antibody stated or its antibody fragment measure the UCH-L1 level in subject to determine whether subject suffers from slightly Traumatic brain injury or in develop mild trauma cerebral injury risk in.Therefore, in certain embodiments, the disclosure is also It provides a kind of for determining whether the subject in the risk with traumatic brain injury or in traumatic brain injury is to use In therapy or the method for the candidate for the treatment of.In general, subject is at least one of following: (i) is lived through to head Damage；(ii) take in and/or be exposed to one or more chemical substances and/or toxin；(iii) suffer from autoimmune disease, Metabolic disorder, brain tumor, anoxic, one or more viruses, meningitis, hydrocephalus suffer from any combination thereof；Or (iv) (i)- (iii) any combination；It (such as suffers from and itself exempts from alternatively, being actually diagnosed in TBI or risk in TBI The subject of epidemic disease, metabolic disorder, brain tumor, anoxic, one or more viruses, meningitis, hydrocephalus or combinations thereof) and/ Or the concentration or amount of unfavorable (i.e. clinically undesirable) UCH-L1 or UCH-L1 segment are shown, as described herein.

A, test sample or biological sample

As used herein, " sample ", " test sample ", " biological sample " refer to containing or the doubtful fluid containing UCH-L1 Sample.Sample can be originated from any suitable source.In some cases, sample may include the fine-particle solid of liquid, flowing Or the fluid suspension of solid particle.In some cases, sample can be handled before analysis described herein.For example, can Before analysis by sample from its source isolated or purified；However, in some embodiments, can directly measure containing The untreated samples of UCH-L1.In a specific example, the source of UCH-L1 is that (such as body fluid, blood are (such as by body material Whole blood, serum, blood plasma), urine, saliva, sweat, sputum, sperm, mucus, tear, lymph, amniotic fluid, interstitial fluid, lung lavage Liquid, celiolymph, excrement, tissue, organ etc.).Tissue may include but be not limited to skeletal muscle tissue, hepatic tissue, lung tissue, kidney group It knits, cardiac muscular tissue, brain tissue, marrow, cervical tissue, skin etc..Sample can be the liquid of fluid sample or solid sample Extract.In some cases, the source of sample can be organ or tissue, such as biopsy samples, can pass through tissue point Solution/cell cracking and dissolve.

It can analyze the fluid sample of wide scope volume.In some illustrative embodiments, sample volume be can be about 0.5nL, about 1nL, about 3nL, about 0.01 μ L, about 0.1 μ L, about 1 μ L, about 5 μ L, about 10 μ L, about 100 μ L, about 1mL, about 5mL or about 10mL etc..In some cases, the volume of fluid sample between about 0.01 μ L and about 10mL, about 0.01 μ L and about 1mL it Between, in about 0.01 μ L and about 100 μ L or between about 0.1 μ L and about 10 μ L.

In some cases, fluid sample can be diluted before in measuring.For example, in the source of UCH-L1 In embodiment for human body fluid (such as blood, serum), fluid can (such as buffer, such as PBS be slow with solvent appropriate Fliud flushing) it is diluted.Fluid sample can be diluted into about 1 times, about 2 times, about 3 times, about 4 times, about 5 times, about 6 before the use Again, about 10 times, about 100 times or more.In other cases, fluid sample is before in measuring without dilution.

In some cases, sample can be with undergoing analysis pre-treatment.Analysis pre-treatment can provide additional functionality, all Such as nonspecific protein removal and/or mixed function that is effective but can inexpensively realizing.The conventional method for analyzing pre-treatment can To include using electric power capture, AC electrodynamics, surface acoustic wave, isotachophoresis, dielectrophoresis, electrophoresis or known in the art Other pre-concentration technologies.In some cases, fluid sample can be concentrated before in measuring.For example, in UCH- The source of L1 is that can pass through precipitating, evaporation, filtering, centrifugation or its group in the embodiment of human body fluid (such as blood, serum) It closes and carrys out concentrating streams.Before the use can by fluid sample be concentrated about 1 times, about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, About 10 times, about 100 times or more.

B, it compares

It may want to include control sample.Control sample can be with the sample same time-division as described above from subject Analysis.The result obtained from Samples subjects can be compared with the result obtained from control sample.Standard song can be provided The measurement result of sample can be compared by line with it.If such standard curve is presented as survey using fluorescent marker Order position, the i.e. marker of the function of fluorescence signal intensity are horizontal.Using the sample for being derived from multiple donors, can provide for just The reference levels of UCH-L1 in normal health tissues, and for obtained from the confession that may have one or more features described above The standard curve of " risky " level of UCH-L1 in the tissue of body.

Therefore, it in view of above, provides a kind of for determining the side of the presence of UCH-L1 in test sample, amount or concentration Method.The method includes measuring the UCH-L1 of test sample by immunoassay, the immunoassay is for example, by using at least A kind of be different from capture antibody epitope on the capture antibody and at least one combination UCH-L1 of epitope on combination UCH-L1 Epitope and optionally include detectable label detection antibody, and including depositing for UCH-L1 in test sample will be used as It is deposited in the signal generated by detectable label of, amount or concentration directly or indirectly indicated with as UCH-L1 in caliberator It is compared in, amount or the generation signal directly or indirectly indicated of concentration.Caliberator is optionally and preferably a series of schools A part of quasi- object, wherein every kind of caliberator is different from other caliberators in series for UCH-L1 concentration.

14, kit

There is provided herein a kind of kit, the kit can be used for measuring or assessing the UCH-L1 or UCH- of test sample L1 segment.Kit includes at least one component for measuring the UCH-L1 of test sample, for measuring test sample The specification of UCH-L1.For example, kit may include for by immunoassay, such as chemiluminescence particle immunoassays Method measures the specification of the UCH-L1 of test sample.Included specification Pasting is on packaging material or can making in kit It is included for package insert.Although specification is usually the material write or printed, they are not limited to such.The disclosure Cover any medium that can be stored this class declaration and be conveyed to end user.Such medium includes but is not limited to electricity Sub- storage medium (such as disk, tape, magnetic disk cartridge, chip), optical medium (such as CD ROM) etc..As used herein, term " specification " may include providing the address of the internet site of specification.

At least one component may include at least one composition, and it includes the one or more of specific binding UCH-L1 Isolated antibody or its antibody fragment.Antibody can be UCH-L1 capture antibody and/or UCH-L1 detection antibody.

Optionally or additionally, kit may include the caliberator for being measured method or control, for example, purifying and The UCH-L1 being optionally lyophilized, and/or at least one container (such as pipe, microtiter plate or item, it can be with anti-UCH-L1 Dan Ke Grand antibody coating) and/or buffer, buffer or washing buffer are such as measured, any one of them all can provide molten to be concentrated The substrate solution or terminate liquid of liquid, detectable label (such as enzyme label).Preferably, kit must including being measured method The all components needed, i.e. reagent, reference substance, buffer, diluent etc..Specification can also include for generating standard curve Specification.

Kit may further include the reference standard for quantitative UCH-L1.It can be built using reference standard Found the standard curve for interpolation and/or UCH-L1 concentration of extrapolating.Reference standard may include high UCH-L1 concentration level, example Such as from about 100000pg/mL, about 125000pg/mL, about 150000pg/mL, about 175000pg/mL, about 200000pg/mL, about 225000pg/mL, about 250000pg/mL, about 275000pg/mL or about 300000pg/mL；Medium UCH-L1 concentration level, example Such as from about 25000pg/mL, about 40000pg/mL, about 45000pg/mL, about 50000pg/mL, about 55000pg/mL, about 60000pg/ ML, about 75000pg/mL or about 100000pg/mL；And/or low UCH-L1 concentration level, for example, about 1pg/mL, about 5pg/mL, about 10pg/mL, about 12.5pg/mL, about 15pg/mL, about 20pg/mL, about 25pg/mL, about 30pg/mL, about 35pg/mL, about 40pg/ ML, about 45pg/mL, about 50pg/mL, about 55pg/mL, about 60pg/mL, about 65pg/mL, about 70pg/mL, about 75pg/mL, about 80pg/mL, about 85pg/mL, about 90pg/mL, about 95pg/mL or about 100pg/mL.

Any antibody provided in kit, recombinant antibodies such as special to UCH-L1, can mix detectable label, Fluorogen, radioactive segment, enzyme, biotin/avidin protein tag, chromophore, chemiluminescent labeling etc., Huo Zheshi Agent box may include reagent (such as detection antibody) for the reagent of labelled antibody or for detecting antibody and/or for marking The reagent of analyte (such as UCH-L1) or reagent for testing and analyzing object (such as UCH-L1).Antibody, caliberator and/or right According to may be provided in autonomous container or predistribution is into appropriate determination form, such as predistribution is into microtiter plate,

Optionally, kit includes quality control component (such as sensibility group, caliberator and positive control).Quality control Reagent is prepared as well known in the art and is described on the inset of various Immunoassay Products.Sensibility group membership is optionally It for establishing measuring method performance characteristic, and is optionally further the integrality and measuring method of immunoassay kit reagent Standardized available index.

Kit also optionally includes carrying out diagnostic assay method or being conducive to other reagents needed for quality control evaluation, Buffer, salt, enzyme, enzyme cofactor, substrate, detection reagent etc..For separating and/or handling other components of test sample (such as buffer and solution) (such as pretreating reagent) may also comprise in kit.Kit can also comprise one or more A other controls.One or more components of kit can be lyophilized, and kit can further comprise being suitable in said case Restore the reagent of freeze-dried component.

The various components of kit are optionally according to needing to be provided in fitted vessel, such as microtiter plate.Kit Can further comprise for accommodate or the container of stored sample (such as urine, whole blood, blood plasma or blood serum sample container or Box).Where appropriate, kit optionally containing reaction vessels, mixer and be conducive to reagent preparation or test sample its Its component.Kit may also comprise be used to help obtain test sample one or more instruments, such as syringe, pipette, Pliers, measurement spoon etc..

If detectable label is at least one acridine compounds, kit may include at least one acridine -9- formyl Amine, at least one acridine -9- aryl ester carboxylic acid or any combination thereof.If detectable label is at least one acridine compounds, Then kit may also comprise hydrogen peroxide sources, such as buffer, solution and/or at least one alkaline solution.If desired, examination Agent box can contain solid phase, such as magnetic-particle, bead, test tube, microtiter plate, colorimetric cylinder, film, scaffold molecule, film, filter Paper, disc or chip.

If desired, kit may further include one kind for measuring another analyte in test sample or Various ingredients individually or with specification are further combined, and the another kind analyte can be biomarker, such as wound The biomarker of property cerebral injury or illness.

A, kit and adaptation of methods

Kit (or its component), and for being assessed or being measured in test sample by immunoassay as described herein It (includes that of particle including wherein solid phase that the method for the concentration of UCH-L1, which can be adapted for a variety of automated and semi-automatic systems, A bit), such as U.S. Patent number 5,063,081, U.S. Patent Application Publication No. 2003/0170881,2004/0018577,2005/ Described in 0054078 and 2006/0160164 and for example by Abbott Laboratories (Abbott Park, IL) conduct Abbott point-of-care (i-Or i-STAT Alinity, Abbott Laboratories) commercially commercially available, and Those of described in U.S. Patent number 5,089,424 and 5,006,309 and for example by Abbott Laboratories (Abbott Park, IL) conductOr Abbott Alinity device series business is commercially available.

Automation or some differences between automanual system and non-automated system (such as ELISA) include first special (it can influence sandwich formation and analyte to substrate attached by specific binding partner (such as analyte antibody or capture antibody) Reactivity), and length and the time of capture, detection and/or any optionally washing step.Automation or semi-automatic form (such asWhen can have relatively short incubation with any subsequent platform, Abbott Laboratories) Between (such asIt is about 18 minutes), and non-automated form (such as ELISA) about sample and is caught Relatively long incubation time (for example, about 2 hours) may be needed by obtaining reagent.Similarly, automation or semi-automatic form (such asWith any subsequent platform) can have relatively short incubation time (such as It is about 4 minutes), and non-automated form (such as ELISA) may be incubated for relatively long incubation time (for example, about 2 hours) It detects antibody (such as conjugation reagents).

The other platforms that can be obtained from Abbott Laboratories include but is not limited to(ginseng See such as U.S. Patent number 5,294,404, be incorporated hereby accordingly),EIA (bead) and Quantum^TMII and other platforms.In addition, measuring method, kit and kit components can in other forms, such as It is used in electrochemistry or other handheld or point-of-care type measurement system.As previously mentioned, the disclosure for example suitable for Carry out the business Abbott point-of-care (i- of sandwich immunoassaysAbbott Laboratories) electrochemistry exempts from Epidemic disease measures system.Immunosensor and its manufacturing method and the method operated in test device is intended for single use are described in for example beautiful State's patent No. 5,063,081,2003/0170881,2004/0018577,2005/0054078 and of U.S. Patent Application Publication No. In 2006/0160164, the patent is incorporated hereby about introduction in this respect.

Specifically, about measuring method to i-The adaptability of system, preferably following configuration.By the silicon core of micro production Piece manufacture has a pair of golden ampere meter working electrode and silver-silver chloride reference electrode.On a working electrode, there will be immobilization Capture antibody polystyrene bead (diameter 0.2mm) adhere on electrode patterning polyvinyl alcohol polymer coating On.The chip is assembled into the i- with the fluid form suitable for immunoassayBox.In a part of silicon chip On, there are the specific binding partner of UCH-L1, such as one or more UCH-L1 antibody are (one or more to combine Monoclonal/polyclonal antibody of UCH-L1 or the segment of its segment, its variant or its variant) or one or more anti-UCH-L1 The variant of DVD-Ig (or can be in conjunction with its segment, segment of its variant or its variant of UCH-L1), any one of them are ok It is detectably labeled.It is containing the phosphatic aqueous reagent of para-aminophenol in the fluid pouch of box.

In operation, in the holding room for the sample from the doubtful subject with TBI being added to testing cassete, and will Box is inserted into i-In reader.Sample is pushed into the pipeline containing chip by the pump element in box.Make sample and passes Sensor contact, so that enzyme conjugate be made to be dissolved into sample.Sample is vibrated on a sensor, to promote about 2-12 minutes Interlayer is formed.In the penultimate stride of measuring method, sample is pushed into waste compartment and using containing for alkaline phosphatase The wash fluid of the substrate of enzyme come wash excessive enzyme conjugate and from sensor core on piece sample.In the final step of measurement In, alkali phosphatase enzyme mark is reacted with para-aminophenol phosphate to crack bound phosphate groups and allow the para-aminophenol of release It is electrochemically oxidized at working electrode.Based on measured electric current, reader can be determined by embedded mobile GIS and manufactory Calibration curve calculate sample in UCH-L1 amount.

Method as described herein and kit necessarily cover other reagents and method for carrying out immunoassay.Example Such as, cover various buffers, it is all as known in the art and/or to can be readily made or optimize for example for washing, as sewing Close object diluent, particle diluent and/or the buffer as caliberator diluent.Exemplary Conjugate Diluent is certain Employed in kit (Abbott Laboratories, Abbott Park, IL) and contain 2- (N- morpholinyl) ethane sulfonic acid (MES), salt, protein sealer, antimicrobial and detergentConjugate Diluent.It is exemplary Caliberator diluent is employed in certain kits (Abbott Laboratories, Abbott Park, IL)People's caliberator diluent comprising contain MES, other salt, protein sealer and antimicrobial Buffer.In addition, as the U.S. Patent Application No. 61/142 submitted on December 31st, 2008 can be for example in i- described in 048In box-like formula, uses the nucleic acid sequence connecting with signal antibody to obtain improved signal as signal amplification agent and generate. For multiplexing, such as the adaptability of the box of i-Stat in patent document, such as U.S. Patent number 6,438,498 In be described, disclosure is hereby incorporated herein by.

Method as described herein and kit can also relate to monomolecular counting.In some embodiments, for dividing The method of analysis object analysis can be related to assessing analyte present in sample.In some embodiments, assessment can be used for determining The presence and/or concentration of analyte in sample.In some embodiments, the method can also be used to determine present in sample The presence and/or concentration of a variety of difference analytes.

Any monomolecular device for allowing to detect one or more analytes of interest analytes as known in the art can be used for In system as described herein.For example, described device can be microfluidic device, digital micro-fluid device (DMF), based on surface sound The microfluidic device (SAW) of wave, integrated DMF and analyte detection apparatus, integrated SAW and analyte detection apparatus or base In the measurement processing unit of robot.The example for the other devices that can be used includes Quanterix SIMOA^TM Monomolecular counting (the SMC of (Lexington, MA), Singulex^TM) technology (Alameda, CA, see, for example, U.S. Patent number 9, 239,284, content is hereby incorporated herein by) etc..

Although the certain embodiments of this paper are advantageous when being used for and assessing disease (such as traumatic brain injury), Measuring method and kit may be also optionally used for assessing the UCH-L1 in Other diseases, illness and symptom in appropriate circumstances.

Measuring method, which can also be used for identification, improves disease, the compound of such as traumatic brain injury.For example, can make to express The cell of UCH-L1 is contacted with candidate compound.It can be used measuring method as described herein will be in the cell that contacted with compound The expression of UCH-L1 is compared with the expression in control cell.

The present invention has many aspects, and the multiple aspect is illustrated by following non-limiting embodiment.

15, example

It will be apparent to one skilled in the art that method of disclosure as described herein it is other it is suitable modification and change Type is to be easy application and understand, and can make in the case where not departing from the range of the disclosure or aspect disclosed herein and embodiment It is carried out with suitable equivalent.The disclosure has been described in detail, will have to it by reference to following embodiment and more be expressly understood that, institute Some aspects and embodiment that embodiment is intended only to illustrate the disclosure are stated, and are not construed as limiting the scope of the present disclosure.This All journal references, United States Patent (USP) and the disclosed disclosure referred in text is incorporated by by reference accordingly.

Embodiment 1

i-UCH-L1 measuring method

By i-UCH-L1 measuring method is in TBI PATIENT POPULATION research.Using monoclonal antibody to (such as anti- Body A) as capture monoclonal antibody, and use antibody B and C as detection monoclonal antibody.Antibody A is in Abbott The exemplary anti-UCH-L1 antibody developed inside Laboratories (Abbott Park, IL).Antibody B and C identify UCH-L1's Different epitopes and the detection for enhancing antigen in sample are developed by Banyan Biomarkers (Alachua, Floridaa).In The other antibody developed inside Abbott Laboratories (Abbott Park, IL) are being used as capture with various combinations together It also shows that or is expected to show similar signal enhancing when antibody or detection antibody.UCH-L1 is evaluated for key performance attribute Measuring method design.Box configuration is antibody configuration: antibody A (capture antibody)/antibody B+C (detection antibody)；Reagent conditions: 0.8% Solid, 125 μ g/mL Fab alkaline phosphatase cluster conjugates；It is printed with sample introduction: UCH-L1 reference substance.Minute is 10- 15min (the analyte capture time with 7-12min).

Embodiment 2

TBI population research (TRACK-TBI)

Traumatic brain injury Study on Transformation and clinical knowledge (TRACK-TBI) research are a huge and complicated projects. Its mechanism and public-private partnership are made of more than 11 clinical websites, 7 cores, share nearly 50 cooperative institutions, company and Charity organization.Earlier T RACK-TBI Primary Study based on the clinical data from three clinical websites helps refine TBI general Data element, and create the prototype of the TBI Information fair for TRACK-TBI research.

Subject group: recruiting 2700 to 3000 TBI patients in total, average to enter 3 distinguished by clinical nursing path In clinical group: 1, the patient (ED) for evaluating and leaving hospital in emergency department；2, it goes into hospital, but does not live in the patient (ADM) of ICU； And 3, live in the patient (ICU) of ICU.The outer wound of cranium is suffered from by other 100 of each clinical group (n=300) but is not suffered from The patient of TBI recruits 3000 patients as control in total.The layering plan is conducive to comparative effectiveness research (CER) analysis, and It and is not that " slight/moderate/severe " TBI is limited by conventional differentiation.Data collection depend on clinical nursing path (ED, ADM, ) and the requirement of each target ICU.It is 3 groups by the triage in every group, the group defines the data model to be collected It encloses.

Control is the adult plastic surgery trauma patient for meeting following standard: 1, for limbs and/or injury of pelvis and/or Simple Injury score≤4 (non-threat to life) of fracture of rib；2, meet it is identical with TBI subject be included in and exclude mark Standard, in addition to the standard for carrying out CT or MRI to doubtful head injury in ED is not suitable for.By about the loss of consciousness (LOC), consciousness Obstacle and posttraumatic amnesia (PTA)/RA access potential control, and TBI is excluded except current damage；3, it is mentioned for each website The plan of the control number provided out according to the age and Sex distribution that are originated from TBI group is supplied；And 4, will control recruit arrive CA- It to carry out follow-up in MRI group, and goes to a doctor, sent at 2 weeks to comprehensive assessment (CA) if being unable to complete MRI.

Subject's qualification: it has recruited and has gone to a doctor in emergency department (ED), according to U.S.'s medical science of recovery therapy meeting (ACRM) etalon There is the adult patient in all age brackets of acute TBI medical history, the brain function physiology for the traumatic induction that wherein patient suffers from Interrupt, such as show as in following >=1: the loss of consciousness (LOC) of any period；For face accident occur before or after Any loss of memory of event (for example, amnesia)；When accident occurs, any change of the state of mind (feels dizzy, loses Direction and/or confusion)；And/or it may be or may not be permanent focal neurologic impairment.The packet of traumatic induction Include head be knocked, head impact object or brain experience acceleration/deceleration movement (such as whiplass) without to head Direct exterior trauma.

Use be included in/exclusion criteria is shown in table 2.

Table 2

Subject is further arranged in three different assessments by each of group (i.e. ED, ADM and ICU) clinical for 3 One in group: briefly assessment (BA group), pressurization assessment (CA) group or comprehensive assessment+MRI (CA+MRI) group.About Milestone plan with 80% follow-up rate, referring to table 3.

Table 3

Briefly assessment (BA) group in total include 1200 subjects, for ED, ADM and ICU group respectively with 400 by Examination person.Following data is had collected for BA group: demography and complete clinical disease course data；On day 1 (after damage < 24 hours) it is directed to the blood drawing of serum, blood plasma, DNA and RNA；The repetition of serum is directed in the 3-6 hour that baseline is collected on day 1 It draws blood (optionally for the website including the component)；What a part as the process of diagnosis and treatment in hospital was acquired since the 1st day faces Bed computed tomography brain scan；And using the NIH TBI-CDE announced on the website NINDS CDE v.2.0 core final result index measurement, lead to Cross the outcome data that the structuring telephone interview at 2 weeks, 3,6 and 12 months is collected.

Pressurization assessment (CA) group in total include 1200 subjects, for ED, ADM and ICU group respectively with 300 by Examination person+100 controls.Following data is had collected for CA group: demography and complete clinical disease course data；ADM and The high density routine clinical data of ICU group；On day 1 (after damage < 24 hours) it is directed to the blood drawing of serum, blood plasma, RNA and DNA； (optionally for the website including the component) is drawn blood in repetition in the 3-6 hour that baseline is collected on day 1 for serum；For ADM and ICU, on day 3 (48-72 hours) and the blood drawing for serum, blood plasma and RNA in (96-120 hours) the 5th day；The 1st It to the 5th day cerebrospinal fluid collect (optionally for the website including the component)；The a part passed through as diagnosis and treatment in hospital is adopted All clinical computed tomography brain scans of collection；At 2 weeks and 6 months for the blood drawing of serum, blood plasma and RNA；And by using NIH TBI-CDEs v.2.0 core, basic and supplement final result index measurement, by face-to-face 2 weeks, 6 months and structuring in 12 months Access and the outcome data collected at 3 months via structuring telephone interview.

Comprehensive assessment+MRI (CA+MRI) group includes 600 subjects in total, is respectively had for ED, ADM and ICU group 200.Following data is had collected for CA+MRI group: demography and complete clinical disease course data；ADM and ICU group High density routine clinical data；On day 1 (after damage < 24 hours) it is directed to the blood drawing of serum, blood plasma, RNA and DNA；The 1st (optionally for the website including the component) is drawn blood for the repetition of serum in the 3-6 hour that its baseline is collected；For ADM and ICU, on day 3 (48-72 hours) and the blood drawing for serum, blood plasma and RNA in (96-120 hours) the 5th day；On day 1 to 5 days cerebrospinal fluid collects (optionally for the website including the component)；Institute as a part acquisition that diagnosis and treatment in hospital are passed through There is clinical head CT scan；At 2 weeks and 6 months for the blood drawing of serum, blood plasma and RNA；It is acquired at 2 weeks and 6 months 3T studies MRI；And by using NIH TBI-CDEs v.2.0 core, basic and supplement final result index measurement, by 2 The outcome data that week, 6 months and structuring in 12 months are accessed face-to-face and collected at 3 months via structuring telephone interview.

After recruitment, data collection starts in hospital.For CA+MRI patient, 14 days+4 in the day from damage 2 weeks MRI It is completed.Corresponding 2 weeks final results are completed for+3 days 2 weeks MRI's.For CA and BA patient, 2 weeks final results in the day from damage 14 It completes within it+4 days.Final result at 3 months is completed for+7 days 90 days in the day from damage.For CA+MRI patient, at 6 months MRI is in the completions in ± 14 days 180 days from day of injury, wherein ± 14 days in 6 months MRI of corresponding 6 months final results.For CA With BA patient, 6 months final results are completed for+14 days 180 days the day from damage.BTACT should be completed (but not in+7 days of final result It is on the same day and no more than 201 days from damage).± 30 days 360 days in the day from damage of final result at 12 months It completes.

UCH-L1 (table 4) is measured in the subgroup of 59 TRACK TBI patients in i-STAT determination form.

Table 4 is according to the Subject characteristics of CT scan and MRI result

* 24 subjects receive MRI.

Continuous variable is rendered as intermediate value [25-75% interquartile-range IQR] and is compared using prestige formula rank sum test

Or it is rendered as average value (+/- SD) and is compared based on data distribution using t inspection.

Classified variable is rendered as number/total (percentage) and is accurately examined using card side or Fei Sheer.

Other than in the blood drawing in cerebral injury 24 hours, every patient has an extensive medical assessment, including Cranial Computed Tomography, Neuropsychiatry test, Glasgow coma score (GCS), and many patients have also carried out follow-up in damage 2 weeks MRI.According to careful standardization blood drawing scheme and processing, by plasma sample equal part to store at -80 DEG C, then thaws and survey Examination.Two parts of each sample are run, it is listing the result is that the average value run twice.Fig. 1 shows that UCH-L1 level is being damaged It is associated with damage in initial 24 hours (ranges about 2-23 hours) after wound.Table 4 show ethyl alcohol (ETOH) it is horizontal not with life Substance markers object level is associated (Pearson came correlation=0.023, p value=0.89), because ETOH consumption is often and TBI, especially It is severe TBI related.

Table 5 shows the possibility final result of the TBI patient using UCH-L1 reference levels confirmed by CT scan.

Table 5

Sensibility is determined by: TP/T_TBIIt is determined by with specificity: TN/T_{Non- TBI}PPV passes through TP/T_{It is positive}Really It is fixed.NPV passes through TN/T_{It is negative}It determines.Accuracy passes through (TP+TN)/T_{All subjects}It determines.

The result of UCH-L1 cutoff level using 100pg/mL and 300pg/mL of the display of table 6 compared with imaging results. Using these UCH-L1 cutoff values, the subject that suffers from or may suffer from head injury can be evaluated with the presence or absence of traumatic Cerebral injury.For example, disclosed method can determine that the subject of the UCH-L1 level at least 100pg/mL is based on imaging As a result the probability for suffering from traumatic brain injury is greater than 86%.In addition, disclosed method, which can determine, has at least 300pg/ The subject of the UCH-L1 level of mL is greater than 96% based on the probability that imaging results suffer from traumatic brain injury.

UCH-L1 measuring method, biomarker knot are used for using the reference levels of 300pg/mL in the sample size group Extension Glasgow final result when fruit height indicates 3 months scores (GOSE) (p value=0.0082).UCH-L1 level is after injury Initial 24 hours (ranges about 2-23 hours) in it is associated with damage.The appearance of marker to disappear time be more than The initial expected of the measuring method.The data illustrate need side of the UCH-L1 measuring method in prediction TBI patient to head CT/MRI Excellent sensibility/the specificity and amplitude in face.

Fig. 2 and Fig. 3 show horizontal compared to the UCH-L1 at time point 1 according to time point (Fig. 2) and according to time point 2 Recipient's operating characteristics (ROC) to the UCH-L1 of all subjects and CT scan result of variation (Fig. 3) analyze.Fig. 4 shows Go out according to absolute magnitude (" absolute increment ") (UCH-L1 level i.e. in first time point (" time point 1 ") to the second time point The variation between UCH-L1 level in (" time point 2 ")) it is horizontal to the UCH-L1 of all subjects and CT scan result Recipient's operating characteristics (ROC) analysis.

Table 7 shows the TRACK data analysis to 191 Patient Sample A's sizes, specifically, to time point 2 and time point 1 Between UCH-L1 level variation analysis.1 sample of time point is being obtained from damage 24 hours, and in 1 sample of time point About 3-6 hours acquirement 2 sample of time point after product obtain.UCH-L1 seems the (n=that has significant change from time point 1 to time point 2 191, intermediate value increment (i.e. from the variation at time point 1 to time point 2)=- 38pg/mL, Wilcoxon symbol rank inspection p value < 0.0001).See also Fig. 5 and Fig. 6.Fig. 5 shows box traction substation, indicates all trouble at time point 1 and time point 2 in research The UCH-L1 measured in person is horizontal.Fig. 6 shows the measurement dynamics intermediate value Z-score according to the test result at time point.These Data only compare the analysis at time point, rather than based on CT, MRI or GCS+/-.

Table 7

Based on when 1 sample of time point is obtained, for example, 0 to about 6 hour (" 0-6 hours groups ") after damage, about 6 after damage To about 12 hours (" 6-12 hours groups ") further analysis the TRACK data from 191 samples and the TRACK data with Cranial Computed Tomography scanning result and/or GCS scoring are associated.By in 2 sample of time point 1 and time point UCH-L1 level and it is positive or Negative Cranial Computed Tomography scanning result and/or instruction is slight or moderate/severe TBI GCS scoring is compared.Since the analysis is based on The time of 1 sample of time point is obtained, therefore every group of subject's quantity is seldom.Referring to table 8.

Table 8

^*The time of the first sample of acquirement from trauma time

CT scan.Fig. 7 A and Fig. 7 B are shown from 0-6 hours groups, the subject with positive or negative head CT scan In UCH-L1 result.Fig. 8 A and Fig. 8 B are shown from 6-12 hours groups, with the tested of positive or negative head CT scan UCH-L1 result in person.Fig. 7 A and Fig. 8 A are shown in the subject with positive or negative CT scan in 1 He of time point The distribution of UCH-L1 level at time point 2.UCH-L1 for 0-6 hours groups and 6-12 hours groups, in CT positive subjects Level is above CT negative subject at time point 1 and time point 2.From 0-6 hours groups, with the tested of positive CT scan UCH-L1 level in person has significant reduce from time point 1 to time point 2.Fig. 7 B and Fig. 8 B are shown 1 He of time point UCH-L1 level ROC curve associated with CT scan result at time point 2.0-6 hours and 6- are directed to based on ROC curve The sensibility and specificity of the reference levels of 12 hours groups is shown in table 9.

Table 9

Time range	Reference levels (pg/mL)	Sensibility	Specificity
				0-6	370	100	37.5
0-6	509	100	75
				6-12	96	96	30
6-12	98	86	35

Fig. 9 A shows 2 sample of time point in 0-6 hours groups, the subject with positive or negative head CT scan Variation (" increment ") of the condition for the UCH-L1 level of 1 sample of time point, and Figure 10 A shows the UCH-L1 level of the group Absolute magnitude (" absolute increment ").Fig. 9 B is shown from 6-12 hours groups, the subject with positive or negative head CT scan In 2 sample of time point relative to 1 sample of time point UCH-L1 level variation (" increment "), and Figure 10 B shows the group UCH-L1 level absolute magnitude (" absolute increment ").Compared with the subject with negative head CT scan, there is positive head The subject's of portion's CT scan has bigger reduction or variation from the UCH-L1 level at time point 1 to time point 2.With come from 6- The subject of 12 hours groups compares, and there is the subject from 0-6 hours groups bigger UCH-L1 level to change.Analyze data with Check wherein acquirement 1 sample of time point (" 0-10 hours groups ") and by itself and corresponding time between 0-10 hour after injury The sample that is compared of 2 samples of point, and wherein more than 10 hours 1 samples of acquirement time point (" > 10 hours groups ") after injury And the sample for being compared it with corresponding 2 sample of time point.Figure 17 and Figure 18 respectively illustrates UCH-L1 is horizontal Absolute magnitude (" absolute increment ") ROC curve associated with 0-10 hours groups and > 10 hours CT scan results of group.Based on ROC The sensitivity of the absolute magnitude (or variation between time point 1 and time point 2) of the UCH-L1 level for 0-10 hours groups of curve Property and specificity are shown in table 10.

Table 10

Time range	Absolute magnitude (pg/mL)	Sensibility (%)	Specific (%)
				0-10	25	85	41
0-10	23	90	35

For the multiple mutation analysis of CT scan.0-6 hours group 12-14 name subject in determination time point 1 and when Between point 2 level between multiple variation.It is horizontal that Figure 16 shows UCH-L1 associated with positive or negative head CT scan Multiple variation (UCH-L1 level at time point 1/UCH-L1 at time point 2 is horizontal) ROC curve.If after injury First 6 hours in obtain 1 sample of time point, then 1.5 multiple reference levels (i.e. 1 result of time point/time point, 2 result) have 83% sensibility and 75% specificity.The multiple reference levels show well in terms of the positive CT in prediction subject, Wherein change less than 1.5 multiple predictive of the positive CT scan result in subject.

The multiple variation between time point 1 and the level at time point 2 is being determined in the subject of 0-12 hours groups.UCH-L1 The ROC curve of horizontal multiple variation (UCH-L1 level/UCH-L1 at time point 2 at time point 1 is horizontal) and it is positive or Negative head CT scan is associated (data are not shown).If obtaining 1 sample of time point in first 12 hours after injury, 1.81 multiple reference levels (i.e. 1 result of time point/time point, 2 result) have 93% sensibility and 36% specificity. The multiple reference levels showed in terms of the positive CT in prediction subject it is good, wherein multiple variation less than 1.81 predictive of Positive CT scan result in subject and therefore subject is classified as to need CT scan.The multiple of UCH-L1 level changes The ROC curve and positive or negative head CT scan of (UCH-L1 level/UCH-L1 at time point 1 at time point 2 is horizontal) Associated (data are not shown).If obtaining 2 sample of time point in first 12 hours after injury, 0.55 multiple refers to water Flat (i.e. 1 result of time point/time point, 1 result) has 93% sensibility and 36% specificity.The multiple reference levels exist Performance is good in terms of predicting the positive CT in subject, wherein be more than 0.55 multiple variation predictive of the positive CT in subject Scanning result and therefore subject is classified as to need CT scan, and subject is classified as pair by the multiple variation less than 0.55 Need CT scan feminine gender.

ROC analysis is carried out also to show if the UCH-L1 level at time point 1 is greater than 550pg/mL or if time Multiple variation between point 2 and time point 1 is greater than 0.55, then is classified as subject positive to CT scan is needed；If when Between put that UCH-L1 level at 1 is less than 550pg/mL or if the multiple variation between time point 2 and time point 1 is less than 0.55, then subject is classified as negative to CT scan is needed.These reference levels are with 100% sensibility and 32% Specificity.

GCS scoring.Figure 11 A and Figure 11 B show be accredited as from 0-6 hours groups, based on GCS scoring with slight or UCH-L1 result of the moderate into the subject of severe (" moderate/severe ") TBI.Figure 12 A and Figure 12 B show small from 6-12 When group, be accredited as based on GCS scoring UCH- with slight or moderate into the subject of severe (" moderate/severe ") TBI L1 result.Figure 11 A and Figure 12 A show be confirmed as in slight or moderate/severe subject time point 1 and when Between put the distribution of UCH-L1 level at 2.UCH-L1 for 0-6 hours groups and 6-12 hours groups, in moderate/severe subject Level is all remarkably higher than slight subject at time point 1 and time point 2.UCH-L1 in 0-6 hours groups and 6-12 hours groups Level has significant reduce from time point 1 to time point 2.Figure 11 B and Figure 12 B are shown will be at time point 1 and time point 2 UCH-L1 level ROC curve associated with GCS scoring.The reference levels of the group of 0-6 hour and 6-12 hours based on ROC curve Sensibility and specificity be shown in table 11.

Table 11

Figure 13 A shows UCH-L1 water of 2 sample of time point relative to 1 sample of time point in the subject of 0-6 hours groups Flat variation (" increment ").Figure 13 B show 6-12 hours group subject in 2 sample of time point relative to 1 sample of time point UCH-L1 level variation (" increment ").

Figure 14 A shows absolute magnitude (or the change between time point 1 and time point 2 of the UCH-L1 level of 0-6 hours groups Change；" absolute increment ") and Figure 14 B show time point 1 and UCH-L1 level at time point 2 and GCS score (slight phase For moderate/severe) associated ROC curve.Figure 15 A show 6-12 hours group UCH-L1 level absolute magnitude (or when Between point 1 and time point 2 between variation；(" absolute increment ") and Figure 15 B are shown will be at time point 1 and time point 2 UCH-L1 level ROC curve associated with GCS score (slightly relative to moderate/severe).It is determined suffering from based on GCS scoring There is the subject of slight TBI to compare, based on GCS scoring be determined with moderate/severe TBI subject from time point 1 to The UCH-L1 level at time point 2 has bigger reduction or variation.It is small from 0-6 compared with the subject from 6-12 hours groups When group moderate/severe subject there is significant bigger UCH-L1 level variation.Based on ROC curve for 0-6 hour with The sensibility and specificity of the absolute magnitude (or variation between time point 1 and time point 2) of the UCH-L1 level of 6-12 hours groups It is shown in table 12.

Table 12

Time range	Absolute magnitude (pg/mL)	Sensibility	Specificity
				0-6	2528	100	100
0-12	129	70	92

Analyze data to check sample, wherein 0-10 hours after injury (" 0-10 hours groups "), 0-11 is small after injury When (" 0-11 hours groups) " or 0-12 hours after injury (" 0-12 hours groups) " between obtain 1 sample of time point, and by its It is compared with corresponding 2 sample of time point.Data are analyzed to check sample, wherein more than the 11 hours acquirement time after injury Point 1 sample (" > 11 hours groups "), and it is compared with corresponding 2 sample of time point.Figure 19 and Figure 20 are respectively illustrated The absolute magnitude (" absolute increment ") of UCH-L1 level is associated with 0-11 hours groups and > 11 hours CT scan results of group ROC curve.Figure 21 and Figure 22 is respectively illustrated the absolute magnitude (" absolute increment ") of UCH-L1 level and 0-10 hours groups and 0- The associated ROC curve of CT scan result of 12 hours groups.Based on ROC curve for 0-10 hours, 0-11 and 0-12 group The sensibility and specificity of the absolute magnitude (or variation between time point 1 and time point 2) of UCH-L1 level is shown in table 13.

Table 13

Time range	Absolute magnitude (pg/mL)	Sensibility (%)	Specific (%)
				0-10	25	100	36
0-11	25	100	32
				0-12	129	75	76

For the multiple mutation analysis of GCS scoring.Time point 2 and time point 1 are determined in the subject of 0-12 hours groups Level between multiple variation.Multiple variation (the UCH-L1 level at time point 2/at time point 1 of UCH-L1 level UCH-L1 is horizontal) ROC curve it is associated with GCS scoring (data are not shown).If obtained in first 12 hours after injury 2 sample of time point, then 0.73 multiple reference levels (i.e. 1 result of time point/time point, 1 result) with 67% sensibility and 48% specificity.The multiple reference levels show in terms of the moderate to severe TBI in prediction subject well, wherein being greater than Or the multiple equal to 0.73 changes predictive of the moderate in subject to severe TBI.

ROC analysis is carried out also to show if UCH-L1 level at time point 1 is greater than or equal to 550pg/mL or such as Multiple variation between fruit time point 2 and time point 1 is greater than 0.73, then subject is classified as with moderate to severe TBI；Such as UCH-L1 level at fruit time point 1 is less than 550pg/mL or if the multiple variation between time point 2 and time point 1 is less than 0.73, then subject is classified as with slight TBI.These reference levels with 100% sensibility and 42.5% it is special Property.

ROC analysis is carried out also to show if UCH-L1 level at time point 1 is greater than or equal to 450pg/mL or such as Multiple variation between fruit time point 2 and time point 1 is greater than 0.73, then subject is classified as with moderate to severe TBI；Such as UCH-L1 level at fruit time point 1 is less than 450pg/mL or if the multiple variation between time point 2 and time point 1 is less than 0.73, then subject is classified as with slight TBI.These reference levels have 100% sensibility and 40% specificity.

ROC analysis is carried out also to show if UCH-L1 level at time point 1 is greater than or equal to 350pg/mL or such as Multiple variation between fruit time point 2 and time point 1 is greater than 0.73, then subject is classified as with moderate to severe TBI；Such as UCH-L1 level at fruit time point 1 is less than 550pg/mL or if the multiple variation between time point 2 and time point 1 is less than 0.73, then subject is classified as with slight TBI.These reference levels with 100% sensibility and 32.5% it is special Property.

In short, present disclose provides the supplements for being used as standard clinical assessment or alternative form to evaluate subject's The specific UCH-L1 reference value of traumatic brain injury and corresponding measuring method.

It should be understood that previously detailed description and accompanying embodiment are merely illustrative, and it is not construed as to the scope of the present invention Limitation, the range only limits by appended claims and its equivalent.

The various changes and modifications of disclosed embodiment it will be apparent to those skilled in the art that.Without departing from Under the premise of its spirit and scope, it can carry out and chemical structure including but not limited to of the invention, substituent group, derivative, centre Body, synthesis, composition, preparation and/or the related such change of application method and modification.

For sake of completeness, various aspects of the invention are listed in following number clause:

Clause 1, a kind of assisted diagnosis and evaluation suffer from or may suffer from the human experimenter's of the damage to head Method, which comprises a) sample for being derived from the subject in 24 hours after to head suspicious lesion is measured, To measure or detect the level of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in the sample；And b) determine that the subject is It is no to suffer from slight or moderate to severe trauma cerebral injury (TBI), wherein (i) when the UCH-L1 level in the sample is higher than When the reference levels of UCH-L1, the subject is determined as with moderate to severe trauma cerebral injury, and works as the sample When UCH-L1 level in product is lower than the reference levels of UCH-L1, the subject is determined as damaging with mild trauma brain Wound；(ii) when from the UCH-L1 level in the sample that first time point obtains in the sample that the second time point obtained When increasing or decreasing, the subject is determined as with moderate to severe trauma with statistically significant for UCH-L1 level Property cerebral injury, and when from the UCH-L1 level in the sample that first time point obtains to the sample obtained at the second time point In UCH-L1 level it is not statistically significant when increasing or decreasing, the subject is determined as with mild trauma Cerebral injury；(iii) when the level of UCH-L1 decreases or increases at least absolute magnitude to second sample from first sample, The subject is determined as with moderate to severe trauma cerebral injury, or horizontal from first sample as UCH-L1 When not decreasing or increasing at least absolute magnitude to second sample, the subject is determined as damaging with mild trauma brain Wound；(iv) when the level of UCH-L1 decreases or increases at least the first absolute magnitude to second sample from first sample, The subject is determined as with moderate to severe trauma cerebral injury, or horizontal from first sample as UCH-L1 When decreasing or increasing at least the second absolute magnitude to second sample, the subject is determined as damaging with mild trauma brain Wound；Or it ought (v) take when the UCH-L1 level in the sample is higher than the reference levels of UCH-L1 or from first time point The UCH-L1 level in sample obtained dramatically increases or reduces X amount to the UCH-L1 level in the sample that the second time point obtained When, the subject is determined as with moderate to severe trauma cerebral injury, and when the UCH-L1 in the sample is horizontal When reference levels lower than UCH-L1 or when from the UCH-L1 level in the sample that first time point obtains at second Between put in the sample of acquirement UCH-L1 level do not dramatically increase or reduce when being greater than X amount, the subject is determined as suffering from There is mild trauma cerebral injury.

Clause 2, the method as described in clause 1, in which: the first time point is after the suspicious lesion about 0 to about 12 hours, and statistically significant the increasing or decreasing is to be more than from second time point to the first time point About 1 times；The first time point is about 0 to about 12 hour after the suspicious lesion, and the statistically significant increasing Add deduct is from second time point to the first time point less more than about 0.73 times；Or the first time point is institute About 0 to about 12 hour after stating suspicious lesion, the reference levels of the UCH-L1 between about 350pg/mL and about 550pg/mL, And statistically significant the increasing or decreasing is from second time point to the first time point more than about 0.73 Times.

Clause 3, the method as described in clause 1 or 2, wherein the subject receives before or after carrying out the measurement Cross Golmud-Lhasa Pipeline scoring.

Clause 4, the method as described in clause 3, wherein being scored based on the Golmud-Lhasa Pipeline, the subject is doubted Like with moderate to severe trauma cerebral injury.

Clause 5, the method as described in clause 4, wherein the reference levels are damaged with moderate and severe trauma brain The subject of wound is associated.

Clause 6, the method as described in clause 5, wherein being 3- by the reference levels and Glasgow Glasgow coma scale 12 is associated.

Clause 7, the method as described in clause 3, wherein being scored based on the Golmud-Lhasa Pipeline, the subject is doubted Like with mild trauma cerebral injury.

Clause 8, the method as described in clause 7, wherein by the reference levels with mild trauma cerebral injury by Examination person is associated.

Clause 9, the method as described in clause 8, wherein being 13-15 phase by reference levels and Glasgow Glasgow coma scale Association.

Clause 10, the method as described in any one of clause 1 to 9, wherein the reference levels are by having at least about What the measuring method of the specificity between the sensibility and at least about 30% to 100% between 85% to 100% determined.

Clause 11, the method as described in any one of clause 1 to 10, wherein the reference levels are by having at least about 99% sensibility and at least about 75% specificity measuring method determine.

Clause 12, the method as described in any one of clause 1 to 11, wherein the reference levels are at least about 50pg/mL To between about 12000pg/mL.

Clause 13, the method as described in any one of clause 1 to 12, wherein the reference levels are at least about 65pg/mL To between about 9019pg/mL.

Clause 14, the method as described in any one of clause 1 to 13, wherein the sample is 0 to 6 after the suspicious lesion It is obtained in hour and the reference levels is the measurement by having the specificity of about 100% sensibility and at least about 33% What method determined.

Clause 15, the method as described in clause 14, wherein the sample obtains in 0 to 6 hour after the suspicious lesion And the reference levels are about 311pg/mL.

Clause 16, the method as described in any one of clause 1 to 13, wherein the sample is 0 to 6 after the suspicious lesion It is obtained in hour and the reference levels is the measuring method by having the specificity of about 100% sensibility and about 100% Determining.

Clause 17, the method as described in clause 16, wherein the sample obtains in 0 to 6 hour after the suspicious lesion And the reference levels are about 9019pg/mL.

Clause 18, the method as described in any one of clause 1 to 13, wherein the sample is about 6 after the suspicious lesion It is obtained between hour to 12 hours and the reference levels is by having about 100% sensibility and at least about 30% spy What anisotropic measuring method determined.

Clause 19, the method as described in clause 18, wherein the sample is between after the suspicious lesion about 6 to 12 hours It obtains and the reference levels is about 98pg/mL.

Clause 20, the method as described in any one of clause 1 to 13, wherein the sample is about 6 after the suspicious lesion It is obtained between hour to 12 hours and the reference levels is by having about 100% sensibility and at least about 63% spy What anisotropic measuring method determined.

Clause 21, the method as described in clause 20, wherein the sample is between after the suspicious lesion about 6 to 12 hours It obtains and the reference levels is about 209pg/mL.

Clause 22, the method as described in any one of clause 1 to 13, wherein the sample is about 6 after the suspicious lesion It is obtained between hour to 12 hours and the reference levels is by the sensibility and at least about 96% at least about 90% Specificity measuring method determine.

Clause 23, the method as described in clause 22, wherein the sample is between after the suspicious lesion about 6 to 12 hours It obtains and the reference levels is about 569pg/mL.

Clause 24, the method as described in clause 4, wherein by the absolute magnitude and suffering from moderate and severe trauma cerebral injury Subject it is associated.

Clause 25, the method as described in clause 24, wherein being 3- by the absolute magnitude and Glasgow Glasgow coma scale 12 is associated.

Clause 26, the method as described in clause 7, wherein by the absolute magnitude with it is tested with mild trauma cerebral injury Person is associated.

Clause 27, the method as described in clause 26, wherein being 13- by the absolute magnitude and Glasgow Glasgow coma scale 15 is associated.

Clause 28, the method as described in any one of clause 1 to 4,7 or 24 to 27, wherein the absolute magnitude is to pass through tool There is the measuring method of the sensibility between at least about 70% to 100% and the specificity between at least about 30% to 100% to determine.

Clause 29, the method as described in clause 28, wherein the sample obtains in 0 to 6 hour after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of about 100% sensibility and about 100%.

Clause 30, the method as described in clause 29, wherein the sample obtains in 0 to 6 hour after the suspicious lesion And the absolute magnitude is about 2528pg/mL.

Clause 31, the method as described in clause 30, wherein the sample obtains in 6 to 12 hours after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of at least about 70% sensibility and at least about 92%.

Clause 32, the method as described in clause 31, wherein the sample obtains in 6 to 12 hours after the suspicious lesion And the absolute magnitude is about 129pg/mL.

Clause 33, the method as described in clause 28, wherein the sample obtains in 0 to 10 hour after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of about 100% sensibility and at least about 36%.

Clause 34, the method as described in clause 33, wherein the sample obtains in 6 to 12 hours after the suspicious lesion And the absolute magnitude is about 25pg/mL.

Clause 35, the method as described in clause 28, wherein the sample obtains in 0 to 11 hour after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of about 100% sensibility and at least about 32%.

Clause 36, the method as described in clause 35, wherein the sample obtains in 0 to 11 hour after the suspicious lesion And the absolute magnitude is about 25pg/mL.

Clause 37, the method as described in clause 28, wherein the sample obtains in 0 to 12 hour after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of at least about 75% sensibility and at least about 76%.

Clause 38, the method as described in clause 37, wherein the sample obtains in 0 to 12 hour after the suspicious lesion And the absolute magnitude is about 129pg/mL.

Clause 39, a kind of assist in are suffered from or may be suffered from the doubtful human experimenter to the damage on head Traumatic brain injury degree method, which comprises a) at least two samples that obtain from the subject are carried out Measurement, first sample is derived from the subject in 24 hours of the suspicious lesion, and second sample is taking The subject is derived within about 3 to about 6 hours after obtaining first sample；B) traumatic brain is detected at least two sample The early stage biomarker of damage, the early stage biomarker are made of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1), The presence of middle UCH-L1 starts to present in about 0 to about 6 hour after the suspicious lesion；C) first sample and described is measured Second sample respectively in UCH-L1 it is horizontal, and determine that the level of UCH-L1 is from first sample to second sample It reduces or increases；And d) level based on UCH-L1 is to reduce, increase also from first sample to second sample It is to maintain identical, determines the traumatic brain injury degree in the subject.

Clause 40, the method as described in clause 39, wherein first sample is 0 to about 6 hour after the suspicious lesion Interior acquirement.

Clause 41, the method as described in clause 39 or 40, wherein horizontal from first sample to described as UCH-L1 When second sample increases or decreases at least about 20pg/mL at least about 6100pg/mL, determine the subject with moderate to weight Spend traumatic brain injury.

Clause 42, the method as described in clause 41, wherein first sample is 0 to about 6 hour after the suspicious lesion The level of interior acquirement and UCH-L1 increase or decrease at least about 2528pg/mL

Clause 43, the method as described in clause 41, wherein first sample is 6 to about 12 hours after the suspicious lesion The level of interior acquirement and UCH-L1 increase or decrease at least about 129pg/mL

Clause 44, the method as described in clause 41, wherein first sample is 0 to about 10 or about after the suspicious lesion It is obtained in 11 hours and the level of UCH-L1 increases or decreases at least about 25pg/mL.

Clause 45, the method as described in clause 41, wherein first sample is 0 to about 12 hour after the suspicious lesion The level of interior acquirement and UCH-L1 increase or decrease at least about 129pg/mL.

Clause 46, the method as described in any one of clause 1 to 45, the method further includes with traumatic brain injury Therapy treatment is assessed as the human experimenter with moderate to severe trauma cerebral injury.

Clause 47, the method as described in any one of clause 1 to 45, the method further includes monitorings to be assessed as suffering from There is the human experimenter of mild trauma cerebral injury.

Clause 48, it is a kind of assist in determining whether to suffer from or may suffer from the mankind of the doubtful damage to head by The method that examination person carries out head computerized tomography (CT) scanning, which comprises a) obtained at least to from the subject Two samples are measured, and first sample is derived from the subject in 24 hours of the suspicious lesion, and described Second sample is derived from the subject in about 3 to about 6 hours after obtaining first sample；B) at least two sample The early stage biomarker of traumatic brain injury is detected, the early stage biomarker is by ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) it forms, wherein the presence of UCH-L1 starts to present after the suspicious lesion in about 0 to about 6 hour；C) measurement institute State the first sample and second sample respectively in UCH-L1 it is horizontal, and determine that UCH-L1's is horizontal from first sample It is to reduce or increase to second sample；And it is d) horizontal from first sample to second sample based on UCH-L1 Product be reduce, increase also be to maintain it is identical, it is determined whether to the subject carry out CT scan.

Clause 49, the method as described in clause 48, in which: (i) is higher than when the UCH-L1 level in first sample CT scan is carried out to the subject when reference levels of UCH-L1, and the UCH-L1 level in described a sample is lower than CT scan is not carried out to subject when the reference levels of UCH-L1；(ii) when from the UCH-L1 level in first sample to institute The UCH-L1 level stated in the second sample carries out CT scan to the subject when increasing or decreasing with statistically significant, And when no statistically aobvious from the UCH-L1 level in first sample to the UCH-L1 level in second sample What is write does not carry out CT scan to the subject when increasing or decreasing；(iii) when UCH-L1 it is horizontal from first sample to CT scan carried out to the subject when second sample decreases or increases at least absolute magnitude, and when UCH-L1 it is horizontal from CT scan is not carried out to the subject when first sample does not decrease or increase at least absolute magnitude to second sample； Or (iv) when the level of the UCH-L1 in first sample is higher than the reference levels of UCH-L1 or works as from first sample UCH-L1 level in product to the UCH-L1 level in second sample have it is statistical dramatically increase or reduce when pair The subject carries out CT scan, and when the level of the UCH-L1 in first sample is lower than the reference levels of UCH-L1 Or when not statistical from the UCH-L1 level in first sample to the UCH-L1 level in second sample CT scan is not carried out to the subject when dramatically increasing or reducing.

Clause 50, the method as described in clause 49, in which: the first time point be after the suspicious lesion about 0 to About 12 hours, and statistically significant the increasing or decreasing is small from the first time point to second time point In about 2 times；The first time point is about 0 to about 12 hour after the suspicious lesion, and described statistically significant Increase or decrease is to be less than about 1.81 times from the first time point to second time point；The first time point is described About 0 to about 12 hour after suspicious lesion, and the statistically significant increase is from second time point to described First time point is more than about 0.50 times；The first time point is about 0 to about 12 hour after the suspicious lesion, and described Statistically significant increase is from second time point to the first time point more than about 0.55 times；Or described first Time point is about 0 to about 12 hour after the suspicious lesion, and the reference levels of the UCH-L1 are about 550pg/mL, and institute Stating statistically significant increase is from second time point to the first time point more than about 0.55 times.

Clause 51, the method as described in any one of clause 48 to 50, wherein the subject is carrying out the measurement It is preceding or received CT scan later.

Clause 52, the method as described in clause 51, wherein being based on the CT scan, the subject is doubtful with traumatic Cerebral injury.

Clause 53, the method as described in any one of clause 49,51 or 52, wherein by the reference levels and positive head Computed tomography is associated.

Clause 54, the method as described in clause 49 or 51 to any one of 53, wherein the reference levels are by having What the measuring method of the specificity between the sensibility and at least about 30% to 100% between at least about 80% to 100% determined.

Clause 55, the method as described in clause 49 or 51 to any one of 54, wherein the reference levels are at least about 50pg/mL is between about 1000pg/mL.

Clause 56, the method as described in clause 49 or 51 to any one of 55, wherein the reference levels are at least about 86pg/mL is between about 700pg/mL.

Clause 57, the method as described in clause 49 or 51 to any one of 56, wherein the sample is in the suspicious lesion Afterwards in 0 to 6 hour obtain and the reference levels be by with about 100% sensibility and at least about 37.5% it is special Property measuring method determine.

Clause 58, the method as described in clause 57, wherein the sample obtains in 0 to 6 hour after the suspicious lesion And the reference levels are about 370pg/mL.

Clause 59, the method as described in clause 49 or 51 to any one of 56, wherein the sample is in the suspicious lesion It is obtained in 0 to 6 hour afterwards and the reference levels is by having about 100% sensibility and at least about 75% specificity Measuring method determine.

Clause 60, the method as described in clause 59, wherein the sample obtains in 0 to 6 hour after the suspicious lesion And the reference levels are about 509pg/mL.

Clause 61, the method as described in clause 49 or 51 to any one of 56, wherein the sample is in the suspicious lesion It is obtained between about 6 hours to 12 hours afterwards and the reference levels is by the sensibility and at least about at least about 96% What the measuring method of 30% specificity determined.

Clause 62, the method as described in clause 61, wherein the sample is between after the suspicious lesion about 6 to 12 hours It obtains and the reference levels is about 96pg/mL.

Clause 63, the method as described in clause 49 or 51 to any one of 56, wherein the sample is in the suspicious lesion It is obtained between about 6 hours to 12 hours afterwards and the reference levels is by the sensibility and at least about at least about 86% What the measuring method of 35% specificity determined.

Clause 64, the method as described in clause 63, wherein the sample is between after the suspicious lesion about 6 to 12 hours It obtains and the reference levels is about 86pg/mL.

Clause 65, the method as described in clause 49 or 51 to any one of 56, wherein the sample is in the suspicious lesion It is obtained between about 0 hour to 12 hours afterwards and the reference levels is by the sensibility and at least about with about 100% What the measuring method of 32% specificity determined.

Clause 66, the method as described in clause 65, wherein the sample is between after the suspicious lesion about 6 to 12 hours It obtains and the reference levels is about 550pg/mL.

Clause 67, the method as described in any one of clause 49,51 or 52, wherein the absolute magnitude and positive head are counted Calculation machine tomoscan is associated.

Clause 68, the method as described in clause 67, wherein the absolute magnitude be by have at least about 80% to 100% it Between sensibility and at least about 30% to 100% between specificity measuring method determine.

Clause 69, the method as described in clause 68, wherein the sample obtains in 0 to 10 hour after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of at least about 85% sensibility and at least about 41%.

Clause 70, the method as described in clause 69, wherein the sample obtains in 0 to 10 hour after the suspicious lesion And the absolute magnitude is about 25pg/mL.

Clause 71, the method as described in clause 68, wherein the sample obtains in 0 to 10 hour after the suspicious lesion And the absolute magnitude is determined by the measuring method with the specificity of at least about 90% sensibility and at least about 35%.

Clause 72, the method as described in clause 71, wherein the sample obtains in 0 to 10 hour after the suspicious lesion And the absolute magnitude is about 23pg/mL.

Clause 73, the method as described in any one of clause 1 to 72, wherein second sample after to head injury about 5 hours between about 16 hours or about 3 hours to about 6 hours be obtained from the human experimenter.

Clause 74, the method as described in any one of clause 1 to 73, wherein second sample after to head injury about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours or about 12 Hour is derived from the human experimenter.

Clause 75, the method as described in any one of clause 1 to 74, the method further includes carrying out to the sample Measurement with measure or detect be not UCH-L1 one or more other biomarkers level.

Clause 76, the method as described in clause 75, wherein the measurement include: I, measurement or detection first sample and II, the level of one of described second sample or a variety of other biomarkers determines one or more other biologies The level of marker from first sample to the reduction and/or increase of second sample, and if III, from described the One or more other biomarker levels in a sample into second sample it is described it is one or more its Its biomarker level is decreased or increased with statistically significant, then the human experimenter is evaluated as with slight Traumatic brain injury.

Clause 77, the method as described in clause 75 or 76, wherein one or more other biomarkers be selected from by Group consisting of: S100 β, neuronspecific enolase (NSE), glial fibrillary acidic protein (UCH-L1), Apo rouge Albumen 1, Tau, C reactive protein (CRP), free brain-derived neurotrophic factor (BDNF), p-Tau, total BDNF, Troponin I (TnI) and combinations thereof.

Clause 78, the method as described in clause 77, wherein one or more other biomarkers are S100 β BDNF Or CRP.

Clause 79, the method as described in any one of clause 1 to 78, the method further includes with mild trauma brain Damaging therapies treatment is assessed as the human experimenter with mild trauma cerebral injury.

Clause 80, the method as described in any one of clause 1 to 79, the method further includes monitorings to be assessed as suffering from There is the human experimenter of mild trauma cerebral injury.

Clause 81, the method as described in any one of clause 1 to 80, wherein the level of measurement UCH-L1 is surveyed by immune Determine method progress.

Clause 82, the method as described in any one of clause 1 to 81, wherein the level of measurement UCH-L1 includes: A, makes institute It states sample simultaneously or is successively contacted in any order with following: (1) antibody is captured, in conjunction in UCH-L1 or UCH-L1 segment Epitope is to form capture antibody-UCH-L1 antigenic compound, and (2) detect antibody, it includes detectable label and combine It is not caught with forming UCH-L1 antigen-detection antibody complex so that being formed by the epitope that the capture antibody combines on UCH-L1 Obtain antibody-UCH-L1 antigen-detection antibody complex and B, based on anti-by the capture antibody-UCH-L1 antigen-detection The signal that the detectable label in nanocrystal composition generates, measures the amount or concentration of UCH-L1 in the sample.

Clause 83, the method as described in any one of clause 1 to 82, wherein the sample is selected from the group being made up of: Whole blood sample, blood serum sample, spinal fluid samples and plasma sample.

Clause 84, the method as described in any one of clause 1 to 83, wherein in the human experimenter by being shaken by body Passivity impact that the external mechanical force or other power for moving, causing closed or open head trauma generate one or many is fallen , the sample is obtained later to the damage on head caused by explosion or shock wave or other types of blunt forces wound.

Clause 85, the method as described in any one of clause 1 to 83, wherein taking in or being exposed in the human experimenter The sample is obtained after the combination of chemical substance, toxin or chemical substance and toxin.

Clause 86, the method as described in clause 85, wherein the chemical substance or toxin are fire, mould, asbestos, desinsection Agent, insecticide, organic solvent, paint, glue, gas, organic metal, drug abuse or one or more combination.

Clause 87, the method as described in any one of clause 1 to 83, wherein the sample from suffer from autoimmune disease, The human experimenter of metabolic disorder, brain tumor, anoxic, virus, meningitis, hydrocephalus or combinations thereof obtains.

Clause 88, the method as described in any one of clause 1 to 83, wherein the method can be in anyone class subject Upper progress, without considering factor in the group being made up of: the clinical condition of human experimenter, human experimenter reality Test room value, human experimenter as suffer from the slight, classification of moderate to severe trauma cerebral injury, human experimenter low or The performance of high UCH-L1 level, and wherein the human experimenter may suffer from any event of the damage to head when Sequence.

Clause 89, a kind of assisted diagnosis and evaluation suffer from or may suffer from head injury (such as traumatic brain injury) Human experimenter method, which comprises

A) sample (such as biological sample) for being derived from the subject in 24 hours after doubtful head injury is carried out Measurement, to measure or detect the level of the ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in the sample；And

B) determine whether the subject suffers from slight or moderate to severe trauma cerebral injury (TBI), in which:

(i) when the UCH-L1 level in the sample is higher than the reference levels of UCH-L1, the subject is determined as With moderate to severe trauma cerebral injury, and when the UCH-L1 level in the sample is lower than the reference levels of UCH-L1 When, the subject is determined as with mild trauma cerebral injury；

(ii) when from the UCH-L1 level in the sample that first time point obtains to the sample obtained at the second time point In UCH-L1 level have statistically significant when increasing or decreasing, the subject is determined as with moderate to severe Traumatic brain injury, and when from the UCH-L1 level in the sample that first time point obtains to obtaining at the second time point UCH-L1 level in sample it is not statistically significant when increasing or decreasing, the subject is determined as with slight wound Wound property cerebral injury；

(iii) when the level of UCH-L1 decreases or increases at least absolute magnitude from first sample to second sample When, the subject is determined as with moderate to severe trauma cerebral injury, or horizontal from described first as UCH-L1 When sample does not decrease or increase at least absolute magnitude to second sample, the subject is determined as with mild trauma Cerebral injury；

(iv) when the level of UCH-L1 decreases or increases at least first absolutely from first sample to second sample When amount, the subject is determined as with moderate to severe trauma cerebral injury, or horizontal from described the as UCH-L1 When a sample decreases or increases at least the second absolute magnitude to second sample, the subject is determined as with mild trauma Property cerebral injury；Or

(v) when the UCH-L1 level in the sample is higher than the reference levels of UCH-L1 or when from first time point UCH-L1 level in the sample of acquirement dramatically increases or reduces X to the UCH-L1 level in the sample that the second time point obtained When amount, the subject is determined as with moderate to severe trauma cerebral injury, and when the UCH-L1 water in the sample When the flat reference levels lower than UCH-L1 or when from the UCH-L1 level in the sample that first time point obtains to second When the UCH-L1 level in sample that time point obtains is not dramatically increased or reduced greater than X amount, the subject is determined as With mild trauma cerebral injury.

Clause 90, a kind of method for the head injury (such as traumatic brain injury) for evaluating human experimenter, the method Include:

A) sample (such as biological sample) for being derived from the subject in 24 hours after doubtful head injury is carried out Measurement, to measure the level of the ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1) in the sample；And

(i) when the UCH-L1 level in the sample is higher than the reference levels of UCH-L1, during the subject suffers from It spends to severe TBI, and wherein when the UCH-L1 level in the sample is lower than the reference levels of UCH-L1, the subject Suffer from slight TBI；

(ii) when from the UCH-L1 level in the sample that first time point obtains to the sample obtained at the second time point In UCH-L1 level have statistically significant when increasing or decreasing, the subject suffers from moderate to severe TBI, and And wherein when from the UCH-L1 level in the sample that first time point obtains in the sample that the second time point obtained UCH-L1 level it is not statistically significant when increasing or decreasing, the subject suffers from slight TBI；

(iii) when the level of UCH-L1 decreases or increases at least absolute magnitude from first sample to second sample When, the subject suffers from moderate to severe TBI, and wherein horizontal from first sample to described the as UCH-L1 When two samples do not decrease or increase at least absolute magnitude, subject suffers from slight TBI；Or

(iv) when the level of UCH-L1 decreases or increases at least first absolutely from first sample to second sample When amount, the subject suffers from moderate to severe TBI, and wherein horizontal from first sample to described as UCH-L1 When second sample decreases or increases at least the second absolute magnitude, the subject suffers from slight TBI.

Clause 91, the method as described in clause 89 or 90, in which:

The first time point is about 0 to about 12 hour after the doubtful head injury, and described statistically aobvious What is write increases or decreases selected from the group being made up of:

(a) it is more than about 1 times from second time point to the first time point；

(b) it is more than about 0.73 times from second time point to the first time point；Or

(c) it is more than about 0.73 times, and the reference of the UCH-L1 from second time point to the first time point Level is between about 350pg/mL and about 550pg/mL.

Clause 92, the method as described in any one of clause 89 to 91, wherein the subject is carrying out the measurement It is preceding or received later Golmud-Lhasa Pipeline scoring.

Clause 93, the method as described in clause 92, wherein scored based on the Golmud-Lhasa Pipeline, the subject The doubtful moderate that suffers from is to severe TBI.

Clause 94, the method as described in any one of clause 89 to 93, wherein by the reference levels and suffering from moderate extremely The subject of severe TBI is associated.

Clause 95, the method as described in clause 94, wherein being by the reference levels and Glasgow Glasgow coma scale 3-12 is associated.

Clause 96, the method as described in clause 92, wherein scored based on the Golmud-Lhasa Pipeline, the subject It is doubtful to suffer from slight TBI.

Clause 97, the method as described in any one of clause 89 to 92, wherein by the reference levels and suffering from slight TBI Subject it is associated.

Clause 98, the method as described in clause 97, wherein being by the reference levels and Glasgow Glasgow coma scale 13-15 is associated.

Clause 99, the method as described in any one of clause 89 to 98, wherein the reference levels (a), which pass through, to be had at least The measuring method of specificity between sensibility and at least about 30% to about 100% between about 85% to about 100% determines；(b) By having at least about 99% sensibility and the measuring method of at least about 75% specificity to determine；(c) at least about 50pg/mL To between about 12000pg/mL；Or (d) at least about 65pg/mL between about 9019pg/mL.

Clause 100, the method as described in any one of clause 89 to 99, wherein the sample (a) is on the doubtful head It is obtained in about 0 to about 6 hour after damage and the reference levels is by the sensibility and at least about 33% with about 100% Specificity measuring method determine；(b) acquirement and the reference in about 0 to about 6 hour after the doubtful head injury Level is determined by the measuring method with the specificity of about 100% sensibility and about 100%；(c) in the doubtful head Portion damage after between about 6 hours to about 12 hours obtain and the reference levels be by with about 100% sensibility and What the measuring method of at least about 30% specificity determined；(d) between after the doubtful head injury about 6 hours to about 12 hours It obtains and the reference levels is determined by the measuring method with the specificity of about 100% sensibility and at least about 63% 's；Or (e) between after the doubtful head injury about 6 hours to about 12 hours obtain and the reference levels be to pass through tool There is the measuring method of at least about 90% sensibility and at least about 96% specificity to determine.

Clause 101, the method as described in any one of clause 89 to 100, wherein the sample (a) is on the doubtful head It is obtained in 0 to about 6 hour after damage and the reference levels is about 311pg/mL；(b) after the doubtful head injury 0 to It is obtained in 6 hours and the reference levels is about 9019pg/mL；(c) about 6 hours to about 12 after the doubtful head injury It is obtained between hour and the reference levels is about 98pg/mL；(d) about 6 hours to about 12 after the doubtful head injury It is obtained between hour and the reference levels is about 209pg/mL；Or (e) after the doubtful head injury about 6 hours to about It is obtained between 12 hours and the reference levels is about 569pg/mL.

Clause 102, the method as described in clause 89 or 90, wherein by the absolute magnitude and with moderate to severe TBI's Subject is associated.

Clause 103, the method as described in clause 102, wherein being by the absolute magnitude and Glasgow Glasgow coma scale 3-12 is associated.

Clause 104, the method as described in clause 89 or 90, wherein by the absolute magnitude and the subject for suffering from slight TBI It is associated.

Clause 105, the method as described in clause 104, wherein being by the absolute magnitude and Glasgow Glasgow coma scale 13-15 is associated.

Clause 106, the method as described in any one of clause 89,90 and 102 to 105, wherein the absolute magnitude is to pass through Measuring method with the specificity between the sensibility and at least about 30% to about 100% between at least about 70% to about 100% Determining.

Clause 107, the method as described in clause 106, wherein the sample (a) after the doubtful head injury about 0 to It is obtained in about 6 hours and the absolute magnitude is the measuring method by having the specificity of about 100% sensibility and about 100% Determining；(b) it is obtained in about 6 to about 12 hours after the doubtful head injury and the absolute magnitude is by having at least About 70% sensibility and at least about 92% specificity measuring method determine；(c) after the doubtful head injury about 0 to It is obtained in about 10 hours and the absolute magnitude is the survey by having the specificity of about 100% sensibility and at least about 36% Determine what method determined；(d) it is obtained in about 0 to about 11 hour after the doubtful head injury and the absolute magnitude is by having About 100% sensibility and at least about 32% specificity measuring method determine；Or (e) after the doubtful head injury about In 0 to about 12 hour obtain and the absolute magnitude be by at least about 75% sensibility and at least about 76% it is special Property measuring method determine.

Clause 108, the method as described in clause 106 or 107, wherein the sample: (a) after the doubtful head injury It is obtained in about 0 to about 6 hour and the absolute magnitude is about 2528pg/mL；(b) after the doubtful head injury about 6 to about It is obtained in 12 hours and the absolute magnitude is about 129pg/mL；(c) after the doubtful head injury in about 6 to about 12 hours It obtains and the absolute magnitude is about 25pg/mL；(d) after the doubtful head injury in 0 to 11 hour obtain and it is described Absolute magnitude is about 25pg/mL；Or (e) after the doubtful head injury in about 0 to about 12 hour obtain and the absolute magnitude It is about 129pg/mL.

Clause 109, it is a kind of assist in suffer from or may suffer from it is traumatic in the human experimenter of head injury The method of the severity of brain injury, which comprises

A) at least two samples obtained from the subject are measured, first sample is in the doubtful head The subject is derived from 24 hours of portion's damage, and second sample is about 3 to about 6 small after obtaining first sample When be derived from the subject；

B) the early stage biomarker of traumatic brain injury, the early stage biology mark are detected at least two sample Note object be made of ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1), wherein the presence of UCH-L1 after the suspicious lesion about 0 to Start to present in about 6 hours；

C) measure first sample and second sample respectively in UCH-L1 it is horizontal, and determine the water of UCH-L1 Flat is to reduce or increase from first sample to second sample；And

D) level based on UCH-L1 is to reduce, increase and be also to maintain phase from first sample to second sample Together, determine the traumatic brain injury degree in the subject, wherein determine traumatic brain injury degree include determine it is described by Whether examination person suffers from slight or moderate to severe trauma cerebral injury.

Clause 110, a kind of method for evaluating traumatic brain injury (TBI) degree in human experimenter, the method packet It includes:

A) it is measured with quantitative from general at least two samples (such as biological sample) that the subject obtains Plain carboxyl-terminal hydrolase L1 (UCH-L1), first sample are derived from described tested in 24 hours of doubtful head injury Person, and second sample is derived from the subject in about 3 to about 6 hours after obtaining first sample；With

B) level based on UCH-L1 is to reduce, increase and be also to maintain phase from first sample to second sample Together, the TBI degree in the subject is determined, wherein determining traumatic brain injury degree includes whether determining the subject By slight or moderate to severe trauma cerebral injury.

Clause 111, the method as described in clause 109 or 110, wherein first sample is after the doubtful head injury It is obtained in about 0 to about 6 hour.

Clause 112, the method as described in any one of clause 109 to 111, wherein horizontal from described first as UCH-L1 When sample increases or decreases at least about 20pg/mL at least about 6100pg/mL to second sample, determine that the subject suffers from There is moderate to severe TBI.

Clause 113, the method as described in clause 109 or 110, wherein first sample (a) damages on the doubtful head It is obtained in about 0 to about 6 hour after wound and the level of UCH-L1 increases or decreases at least about 2528pg/mL；(b) described doubtful It is obtained in about 6 to about 12 hours after head injury and the level of UCH-L1 increases or decreases at least about 129pg/mL；(c) institute It is obtained in about 0 to about 10 hour after stating doubtful head injury and the level of UCH-L1 increases or decreases at least about 25pg/mL； (d) it is obtained in about 0 to about 11 hour after the doubtful head injury and the level of UCH-L1 increases or decreases at least about 25pg/mL；Or (e) horizontal increase of acquirement and UCH-L1 add deduct in about 0 to about 12 hour after the doubtful head injury Few at least about 129pg/mL.

Clause 114, the method as described in any one of clause 89 to 113, the method further includes (a) with being directed to The therapy treatment of TBI is assessed as the human experimenter with slight or moderate to severe TBI, (b) after receiving the treatment Monitor the human experimenter.

Clause 115, the method as described in any one of clause 89 to 113, it is evaluated the method further includes monitoring For the human experimenter with slight or moderate to severe TBI.

Clause 116, one kind assist in determining whether to suffering from or may suffer from head injury (such as traumatic brain damage Wound) human experimenter carry out head computerized tomography (CT) scanning method, which comprises

A) it is measured to detect or measure at least two samples (such as biological sample) obtained from the subject In ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1), first sample is derived from 24 hours of the doubtful head injury The subject, and second sample is derived from the subject in about 3 to about 6 hours after obtaining first sample；

B) measure first sample and second sample respectively in UCH-L1 it is horizontal, and determine the water of UCH-L1 Flat is to reduce or increase from first sample to second sample；And

C) level based on UCH-L1 is to reduce, increase and be also to maintain phase from first sample to second sample Together, it is determined whether CT scan is carried out to the subject, in which:

(i) CT scan is carried out when the UCH-L1 level in first sample is higher than the reference levels of UCH-L1, And wherein when the UCH-L1 level in first sample is lower than the reference levels of UCH-L1 without the CT scan；

(ii) when from the UCH-L1 level in first sample to the UCH-L1 level in second sample have system Meter is learned and carries out the CT scan when significantly increasing or decreasing, and wherein when horizontal from the UCH-L1 in first sample To the UCH-L1 level in second sample it is not statistically significant when increasing or decreasing without the CT scan；Or

(iii) when the level of UCH-L1 decreases or increases at least absolute magnitude to second sample from first sample The CT scan is carried out, and wherein when the level of UCH-L1 does not reduce or increases from first sample to second sample Without the CT scan when adding to few absolute magnitude.

Clause 117, one kind evaluate whether the method for carrying out head computerized tomography (CT) scanning to human experimenter, described Method includes:

A) it is measured with quantitative from general at least two samples (such as biological sample) that the subject obtains Plain carboxyl-terminal hydrolase L1 (UCH-L1), first sample are derived from described tested in about 24 hours of doubtful head injury Person, and second sample is derived from the subject in about 3 to about 6 hours after obtaining first sample；With

B) level based on UCH-L1 is to reduce, increase and be also to maintain phase from first sample to second sample Together, it is determined whether CT scan is carried out to the subject, in which:

Clause 118, the method as described in clause 116 or 117, in which: the first time point is the doubtful head damage About 0 to about 12 hour after wound, and statistically significant the increasing or decreasing is from the first time point to described Second time point was less than about 2 times；The first time point is about 0 to about 12 hour after the doubtful head injury, and institute Stating statistically significant increasing or decreasing is to be less than about 1.81 times from the first time point to second time point；It is described First time point is about 0 to about 12 hour after the doubtful head injury, and the statistically significant increase be from Second time point is more than about 0.50 times to the first time point；After the first time point is the doubtful head injury About 0 to about 12 hour, and the statistically significant increase is from second time point to the first time point More than about 0.50 times；Or the first time point is about 0 to about 12 hour after the doubtful head injury, the UCH-L1 Reference levels be about 550pg/mL, and the statistically significant increase is from second time point to described first Time point is more than about 0.55 times.

Clause 119, the method as described in any one of clause 116 to 118, wherein the subject is carrying out the measurement Before or after received CT scan.

Clause 120, the method as described in clause 119, wherein being based on the CT scan, the subject is doubtful to suffer from wound Property cerebral injury.

Clause 121, the method as described in any one of clause 116 to 120, wherein by the reference levels and positive head CT scan is associated.

Clause 122, the method as described in any one of clause 116 to 121, wherein the reference levels (a) are by having The measuring method of specificity between sensibility and at least about 30% to about 100% between at least about 80% to about 100% determines； (b) at least about 50pg/mL between about 1000pg/mL；Or (c) at least about 86pg/mL between about 700pg/mL.

Clause 123, the method as described in any one of clause 116 to 122, wherein the sample (a) is in the doubtful head It is obtained in about 0 to about 6 hour after portion's damage and the reference levels is by the sensibility and at least about with about 100% What the measuring method of 37.5% specificity determined；(b) after the doubtful head injury in about 0 to about 6 hour obtain and institute Stating reference levels is determined by the measuring method with the specificity of about 100% sensibility and at least about 75%；(c) institute It is obtained between about 6 hours to about 12 hours after stating doubtful head injury and the reference levels is by having at least about 96% Sensibility and at least about 30% specificity measuring method determine；(d) after the doubtful head injury about 6 hours to about Between 12 hours obtain and the reference levels be by at least about 86% sensibility and at least about 35% it is special Property measuring method determine；Or (e) between after the doubtful head injury about 0 hour to about 12 hours obtain and the ginseng The level examined is determined by the measuring method with the specificity of about 100% sensibility and at least about 32%.

Clause 124, the method as described in any one of clause 116 to 123, wherein the sample (a) is in the doubtful head It is obtained in about 0 to about 6 hour after portion's damage and the reference levels is about 370pg/mL；(b) in the doubtful head injury It is obtained in about 0 to about 6 hour afterwards and the reference levels is about 509pg/mL；(c) about 6 small after the doubtful head injury It is obtained between about 12 hours and the reference levels is about 96pg/mL；(d) about 6 small after the doubtful head injury It is obtained between about 12 hours and the reference levels is about 86pg/mL；Or (e) about 6 after the doubtful head injury It is obtained between hour to about 12 hours and the reference levels is about 550pg/mL.

Clause 125, the method as described in clause 116 or 117, wherein the absolute magnitude is related to positive head CT scan Connection.

Clause 126, the method as described in clause 116 or 117, wherein the absolute magnitude be by have at least about 80% to What the measuring method of the specificity between the sensibility and at least about 30% to about 100% between about 100% determined.

Clause 127, the method as described in clause 126, wherein the sample: (a) after the doubtful head injury about 0 to It is obtained in about 10 hours and the absolute magnitude is by having at least about 85% sensibility and at least about 41% specificity Measuring method determine；Or (b) after the doubtful head injury in about 0 to about 10 hour obtain and the absolute magnitude be logical Cross what the measuring method at least about 90% sensibility and at least about 35% specificity determined.

Clause 128, the method as described in clause 126, wherein the sample: (a) after the doubtful head injury about 0 to It is obtained in about 10 hours and the absolute magnitude is about 25pg/mL；Or it is (b) about 0 to about 10 small after the doubtful head injury When it is interior acquirement and the absolute magnitude be about 23pg/mL.

Clause 129, the method as described in any one of clause 89 to 128, wherein second sample is in the doubtful head Portion damage after about 5 hours between about 16 hours or about 3 hours to about 6 hours be obtained from the subject.

Clause 130, the method as described in any one of clause 89 to 128, wherein second sample is in the doubtful head About 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 after portion's damage Hour is derived from the subject in about 12 hours.

Clause 131, the method as described in any one of clause 89 to 130, the method further includes to the sample Be measured be not to measure or detect UCH-L1 one or more other biomarkers level.

Clause 132, the method as described in clause 131, wherein the measurement includes:

(a) measure or detect one of first sample and second sample or a variety of other biomarkers Level,

(b) determine the level of one or more other biomarkers from first sample to second sample Decrease or increase, and

If (c) from one or more other biomarker levels in first sample to second sample One or more other biomarker levels in product have it is statistically significant decrease or increase, then will be described tested Person is evaluated as with slight TBI.

Clause 133, the method as described in clause 131 or 132, wherein one or more other biomarkers are selected from The group being made up of: S100 β, neuronspecific enolase (NSE), glial fibrillary acidic protein (UCH-L1), Apo Lipoprotein 1, Tau, C reactive protein (CRP), free brain-derived neurotrophic factor (BDNF), p-Tau, total BDNF, flesh calcium egg White I (TnI) and combinations thereof.

Clause 134, the method as described in clause 133, wherein one or more other biomarkers be S100 β, BDNF or CRP.

Clause 135, the method as described in any one of clause 89 to 134, wherein the method further includes (a) to use needle Human experimenter with slight or moderate to severe TBI is assessed as to the therapy treatment of TBI, (b) is receiving the treatment After monitor the human experimenter.

Clause 136, the method as described in any one of clause 89 to 135, it is evaluated the method further includes monitoring For the human experimenter with slight or moderate to severe TBI.

Clause 137, the method as described in any one of clause 89 to 136, wherein the level of measurement UCH-L1 includes carrying out Immunoassay.

Clause 138, the method as described in any one of clause 89 to 137, wherein the level of measurement UCH-L1 includes:

(a) make the sample simultaneously or successively contacted in any order with following:

(2) antibody is detected, it includes detectable label and in conjunction with the table not combined by the capture antibody on UCH-L1 Position, to form UCH-L1 antigen-detection antibody complex,

(b) based on raw by the detectable label in the capture antibody-UCH-L1 antigen-detection antibody complex At signal, measure the amount or concentration of UCH-L1 in the sample.

Clause 139, the method as described in any one of clause 89 to 138 are made up of wherein the sample is selected from Group: whole blood sample, blood serum sample, spinal fluid samples and plasma sample.

Clause 140, the method as described in any one of clause 89 to 139, wherein in the subject by being shaken by body Passivity impact that the external mechanical force or other power for moving, causing closed or open head trauma generate one or many is fallen , the sample is obtained after head injury caused by explosion or shock wave or other types of blunt forces wound.

Clause 141, the method as described in any one of clause 89 to 139, wherein taking in or being exposed in the subject The sample is obtained after the combination of chemical substance, toxin or chemical substance and toxin.

Clause 142, the method as described in clause 141, wherein the chemical substance or toxin are fire, mould, asbestos, desinsection Agent, insecticide, organic solvent, paint, glue, gas, organic metal, drug abuse or one or more combination.

Clause 143, the method as described in any one of clause 89 to 139, wherein the sample is from suffering from autoimmunity disease The subject of disease, metabolic disorder, brain tumor, anoxic, virus, meningitis, hydrocephalus or combinations thereof obtains.

Clause 144, the method as described in any one of clause 89 to 139, wherein the method can anyone class by It is carried out on examination person, without considering the factor in the group being made up of: the clinical condition of human experimenter, human experimenter Laboratory evaluation, human experimenter as suffer from classification of the slight or moderate to severe TBI, human experimenter it is low or high The performance of UCH-L1 level and human experimenter may suffer from the timing of any event of head injury.

Clause 145, the method as described in any one of clause 89 to 144, wherein the sample is biological sample.

Clause 146, the method as described in any one of clause 89 to 145, wherein the sample is whole blood sample.

Clause 147, the method as described in any one of clause 89 to 145, wherein the sample is blood serum sample.

Clause 148, the method as described in any one of clause 89 to 145, wherein the sample is plasma sample.

Claims

1. a kind of method for the head injury for evaluating human experimenter, which comprises

A) sample for being derived from the subject in 24 hours after doubtful head injury is measured, to measure the sample The level of middle ubiquitin carboxy terminal hydrolase-l 1 (UCH-L1)；With

(i) when the UCH-L1 level in the sample is higher than the reference levels of UCH-L1, the subject suffers from moderate extremely Severe TBI, and wherein when the UCH-L1 level in the sample is lower than the reference levels of UCH-L1, the subject has met with By slight TBI；

(ii) when from the UCH-L1 level in the sample that first time point obtains in the sample that the second time point obtained UCH-L1 level has statistically significant when increasing or decreasing, and the subject suffers from moderate to severe TBI, and its In when from the UCH-L1 level in the sample that first time point obtains to the UCH-L1 in the sample that the second time point obtained Horizontal not statistically significant when increasing or decreasing, the subject suffers from slight TBI；

(iii) when the level of UCH-L1 decreases or increases at least absolute magnitude to second sample from first sample, institute It states subject and suffers from moderate to severe TBI, and wherein when the level of UCH-L1 is from first sample to second sample When product do not decrease or increase at least absolute magnitude, the subject suffers from slight TBI；Or

(iv) when the level of UCH-L1 decreases or increases at least the first absolute magnitude to second sample from first sample, The subject suffers from moderate to severe TBI, and wherein horizontal from first sample to described second as UCH-L1 When sample decreases or increases at least the second absolute magnitude, the subject suffers from slight TBI.

2. according to the method described in claim 1, wherein:

The first time point is about 0 to about 12 hour after the doubtful head injury, and described statistically significant It increases or decreases selected from the group being made up of:

(c) it is more than about 0.73 times, and the reference levels of the UCH-L1 from second time point to the first time point Between about 350pg/mL and about 550pg/mL.

3. method according to any one of claim 1 to 2, wherein the subject before carrying out the measurement or it It is followed by having received Golmud-Lhasa Pipeline scoring.

4. the subject is doubtful according to the method described in claim 3, wherein being scored based on the Golmud-Lhasa Pipeline With moderate to severe TBI.

5. method according to claim 1 to 4, wherein by the reference levels and with moderate to severe The subject of TBI is associated.

6. according to the method described in claim 5, being wherein 3-12 by the reference levels and Glasgow Glasgow coma scale It is associated.

7. the subject is doubtful according to the method described in claim 3, wherein being scored based on the Golmud-Lhasa Pipeline With slight TBI.

8. according to the method in any one of claims 1 to 3, wherein by the reference levels with slight TBI by Examination person is associated.

9. according to the method described in claim 8, being wherein 13-15 by the reference levels and Glasgow Glasgow coma scale It is associated.

10. method according to any one of claim 1 to 9, wherein the reference levels (a), which pass through, to be had at least about The measuring method of specificity between sensibility and at least about 30% to about 100% between 85% to about 100% determines；(b) lead to The measuring method at least about 99% sensibility and at least about 75% specificity is crossed to determine；(c) at least about 50pg/mL extremely Between about 12000pg/mL；Or (d) at least about 65pg/mL between about 9019pg/mL.

11. method according to any one of claim 1 to 10, wherein the sample (a) is in the doubtful head injury It is obtained in about 0 to about 6 hour afterwards and the reference levels is by having about 100% sensibility and at least about 33% spy What anisotropic measuring method determined；(b) after the doubtful head injury in about 0 to about 6 hour obtain and the reference levels It is to be determined by the measuring method with the specificity of about 100% sensibility and about 100%；(c) it is damaged on the doubtful head It is obtained between about 6 hours to about 12 hours after wound and the reference levels is by the sensibility and at least with about 100% What the measuring method of about 30% specificity determined；(d) it is obtained between after the doubtful head injury about 6 hours to about 12 hours And the reference levels are determined by the measuring method with the specificity of about 100% sensibility and at least about 63%； Or (e) between after the doubtful head injury about 6 hours to about 12 hours obtain and the reference levels be by having At least about 90% sensibility and at least about 96% specificity measuring method determine.

12. method according to any one of claim 1 to 11, wherein the sample (a) is in the doubtful head injury It is obtained in 0 to about 6 hour afterwards and the reference levels is about 311pg/mL；(b) 0 to 6 small after the doubtful head injury When it is interior acquirement and the reference levels be about 9019pg/mL；(c) about 6 hours to about 12 small after the doubtful head injury When between obtain and the reference levels be about 98pg/mL；(d) about 6 hours to about 12 small after the doubtful head injury When between obtain and the reference levels be about 209pg/mL；Or (e) about 6 hours to about 12 after the doubtful head injury It is obtained between hour and the reference levels is about 569pg/mL.

13. according to the method described in claim 1, wherein by the absolute magnitude and subject's phase with moderate to severe TBI Association.

14. according to the method for claim 13, wherein being 3-12 by the absolute magnitude and Glasgow Glasgow coma scale It is associated.

15. according to the method described in claim 1, wherein that the absolute magnitude is associated with the subject of slight TBI.

16. according to the method for claim 15, wherein being 13-15 by the absolute magnitude and Glasgow Glasgow coma scale It is associated.

17. according to claim 1 with method described in any one of 13 to 16, wherein the absolute magnitude be by have at least about What the measuring method of the specificity between the sensibility and at least about 30% to about 100% between 70% to about 100% determined.

18. according to the method for claim 17, wherein the sample (a) is about 0 to about 6 small after the doubtful head injury When it is interior acquirement and the absolute magnitude be by with about 100% sensibility and about 100% specificity measuring method determination 's；(b) it is obtained in about 6 to about 12 hours after the doubtful head injury and the absolute magnitude is by having at least about 70% sensibility and at least about 92% specificity measuring method determine；(c) after the doubtful head injury about 0 to about It is obtained in 10 hours and the absolute magnitude is the measurement by having the specificity of about 100% sensibility and at least about 36% What method determined；(d) it is obtained in about 0 to about 11 hour after the doubtful head injury and the absolute magnitude is by having about 100% sensibility and at least about 32% specificity measuring method determine；Or (e) about 0 after the doubtful head injury In to about 12 hours obtain and the absolute magnitude be by at least about 75% sensibility and at least about 76% it is special Property measuring method determine.

19. method described in 7 or 18 according to claim 1, wherein the sample: (a) after the doubtful head injury about 0 to It is obtained in about 6 hours and the absolute magnitude is about 2528pg/mL；(b) about 6 to about 12 hours after the doubtful head injury It is interior acquirement and the absolute magnitude be about 129pg/mL；(c) it is obtained simultaneously in about 6 to about 12 hours after the doubtful head injury And the absolute magnitude is about 25pg/mL；(d) after the doubtful head injury in 0 to 11 hour obtain and the absolute magnitude It is about 25pg/mL；Or (e) after the doubtful head injury in about 0 to about 12 hour obtain and the absolute magnitude be about 129pg/mL。

20. a kind of method of traumatic brain injury (TBI) degree in evaluation human experimenter, which comprises

A) it is measured with quantitative from the ubiquitin carboxy terminal hydrolase-l 1 at least two samples that the subject obtains (UCH-L1), first sample is derived from the subject, and second sample in 24 hours of doubtful head injury The subject is derived within about 3 to about 6 hours after obtaining first sample；And

B) level based on UCH-L1 from first sample to second sample be reduce, increase also be to maintain it is identical, really TBI degree in the fixed subject, wherein determining that traumatic brain injury degree includes determining whether the subject suffers from Slight or moderate is to severe trauma cerebral injury.

21. according to the method for claim 20, wherein first sample is about 0 to about 6 after the doubtful head injury It is obtained in hour.

22. the method according to claim 20 or 21, wherein horizontal from first sample to described the as UCH-L1 When two samples increase or decrease at least about 20pg/mL at least about 6100pg/mL, determine the subject with moderate to severe TBI。

23. the method according to claim 20 or 21, wherein first sample (a) after the doubtful head injury about It is obtained in 0 to about 6 hour and the level of UCH-L1 increases or decreases at least about 2528pg/mL；(b) it is damaged on the doubtful head It is obtained in about 6 to about 12 hours after wound and the level of UCH-L1 increases or decreases at least about 129pg/mL；(c) described doubtful It is obtained in about 0 to about 10 hour after head injury and the level of UCH-L1 increases or decreases at least about 25pg/mL；(d) institute It is obtained in about 0 to about 11 hour after stating doubtful head injury and the level of UCH-L1 increases or decreases at least about 25pg/mL；Or (e) it is obtained in about 0 to about 12 hour after the doubtful head injury and the level of UCH-L1 increases or decreases at least about 129pg/mL。

24. according to claim 1 to method described in any one of 23, the method further includes with the therapy for being directed to TBI Treatment is assessed as the human experimenter with moderate to severe TBI, and optionally after receiving the treatment described in monitoring Human experimenter.

25. the method further includes monitorings to be assessed as suffering from according to claim 1 to method described in any one of 23 The human experimenter of slight TBI.

26. one kind evaluates whether the method for carrying out head computerized tomography (CT) scanning to human experimenter, which comprises

A) it is measured with quantitative from the ubiquitin carboxy terminal hydrolase-l 1 at least two samples that the subject obtains (UCH-L1), first sample is derived from the subject, and second sample in about 24 hours of doubtful head injury Product are derived from the subject in about 3 to about 6 hours after obtaining first sample；And

B) level based on UCH-L1 from first sample to second sample be reduce, increase also be to maintain it is identical, really It is fixed whether CT scan to be carried out to the subject, in which:

(ii) when from the UCH-L1 level in first sample to the UCH-L1 level in second sample have statistics On carry out the CT scan when significantly increasing or decreasing, and wherein when from the UCH-L1 level in first sample to institute State UCH-L1 level in the second sample it is not statistically significant when increasing or decreasing without the CT scan；Or

(iii) when the when progress that from first sample to second sample decreases or increases at least absolute magnitude of the level of UCH-L1 The CT scan, and wherein when the level of UCH-L1 do not decrease or increase from first sample to second sample to Without the CT scan when few absolute magnitude.

27. according to the method for claim 26, in which:

The first time point be about 0 to about 12 hour after the suspicious lesion and the statistically significant increase or Reduction is to be less than about 2 times from the first time point to second time point；

The first time point be about 0 to about 12 hour after the suspicious lesion and the statistically significant increase or Reduction is to be less than about 1.81 times from the first time point to second time point；

The first time point is about 0 to about 12 hour and the statistically significant increasing after the doubtful head injury Add is from second time point to the first time point more than about 0.50 times；

The first time point is about 0 to about 12 hour and the statistically significant increasing after the doubtful head injury Add is from second time point to the first time point more than about 0.55 times；Or

The first time point is about 0 to about 12 hour after the suspicious lesion, and the reference levels of the UCH-L1 are about Between 550pg/mL, and the statistically significant increase is to be more than from second time point to the first time point About 0.55 times.

28. the method according to claim 26 or 27, wherein the subject connects before or after carrying out the measurement CT scan is received.

29. the subject is doubtful to suffer from traumatic brain according to the method for claim 28, wherein being based on the CT scan Damage.

30. the method according to any one of claim 26 to 29, wherein the reference levels and positive Cranial Computed Tomography are swept It retouches associated.

31. the method according to any one of claim 26 to 30, wherein the reference levels (a), which pass through, to be had at least about The measuring method of specificity between sensibility and at least about 30% to about 100% between 80% to about 100% determines；(b) In At least about 50pg/mL is between about 1000pg/mL；Or (c) at least about 86pg/mL between about 700pg/mL.

32. the method according to any one of claim 26 to 31, wherein the sample (a) is in the doubtful head injury It is obtained in about 0 to about 6 hour afterwards and the reference levels is by the sensibility and at least about 37.5% with about 100% What the measuring method of specificity determined；(b) after the doubtful head injury in about 0 to about 6 hour obtain and it is described refer to water Flat determined by the measuring method with the specificity of about 100% sensibility and at least about 75%；(c) in the doubtful head It is obtained between about 6 hours to about 12 hours after portion's damage and the reference levels is by the sensibility at least about 96% What the measuring method of at least about 30% specificity determined；(d) after the doubtful head injury about 6 hours to about 12 hours it Between obtain and the reference levels are the specific measurements by sensibility at least about 86% and at least about 35% What method determined；Or (e) between after the doubtful head injury about 0 hour to about 12 hours obtain and the reference levels be By having the measuring method of about 100% sensibility and at least about 32% specificity to determine.

33. the method according to any one of claim 26 to 32, wherein the sample (a) is in the doubtful head injury It is obtained in about 0 to about 6 hour afterwards and the reference levels is about 370pg/mL；(b) after the doubtful head injury about 0 to It is obtained in about 6 hours and the reference levels is about 509pg/mL；(c) after the doubtful head injury about 6 hours to about It is obtained between 12 hours and the reference levels is about 96pg/mL；(d) after the doubtful head injury about 6 hours to about It is obtained between 12 hours and the reference levels is about 86pg/mL；Or (e) about 6 hours extremely after the doubtful head injury It is obtained between about 12 hours and the reference levels is about 550pg/mL.

34. the method according to claim 26 or 27, wherein the absolute magnitude is associated with positive head CT scan.

35. the method according to claim 26 or 27, wherein the absolute magnitude is by having at least about 80% to about What the measuring method of the specificity between the sensibility and at least about 30% to about 100% between 100% determined.

36. according to the method for claim 35, wherein the sample: (a) about 0 to about 10 after the doubtful head injury It is obtained in hour and the absolute magnitude is the survey by having the specificity of at least about 85% sensibility and at least about 41% Determine what method determined；Or (b) after the doubtful head injury in about 0 to about 10 hour obtain and the absolute magnitude be to pass through tool There is the measuring method of at least about 90% sensibility and at least about 35% specificity to determine.

37. according to the method for claim 36, wherein the sample: (a) about 0 to about 10 after the doubtful head injury It is obtained in hour and the absolute magnitude is about 25pg/mL；Or (b) after the doubtful head injury in about 0 to about 10 hour It obtains and the absolute magnitude is about 23pg/mL.

38. according to claim 1 to method described in any one of 37, wherein second sample is in the doubtful head injury Afterwards about 5 hours between about 16 hours or about 3 hours to about 6 hours be obtained from the subject.

39. according to claim 1 to method described in any one of 37, wherein second sample is in the doubtful head injury About 3 hours afterwards, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours or It is derived from the subject within about 12 hours.

40. the method further includes surveying to the sample according to claim 1 to method described in any one of 39 It is fixed with measure or detect be not UCH-L1 one or more other biomarkers level.

41. according to the method for claim 40, wherein the measurement includes:

(a) measure or detect the water of one of first sample and second sample or a variety of other biomarkers It is flat,

(b) the horizontal from the first sample subtracting to second sample of one or more other biomarkers is determined Less or increase, and

If (c) from one or more other biomarker levels in first sample into second sample One or more other biomarker levels have it is statistically significant decrease or increase, then by the subject It is evaluated as with slight TBI.

42. the method according to claim 40 or 41, wherein one or more other biomarkers be selected from by with The group of lower composition: S100 β, neuronspecific enolase (NSE), glial fibrillary acidic protein (GFAP), Apo lipoprotein 1, Tau, C reactive protein (CRP), free brain-derived neurotrophic factor (BDNF), p-Tau, total BDNF, Troponin I (TnI) and combinations thereof.

43. according to the method for claim 42, wherein one or more other biomarkers are S100 β, BDNF Or CRP.

44. the method further includes with for slight TBI's according to claim 1 to method described in any one of 43 Therapy treatment is assessed as the subject with slight TBI, and optionally monitored after receiving the treatment mankind by Examination person.

45. the method further includes monitorings to be assessed as suffering from according to claim 1 to method described in any one of 44 The human experimenter of slight TBI.

46. according to claim 1 to method described in any one of 45, wherein the level for measuring the UCH-L1 includes being exempted from Epidemic disease measuring method.

47. according to claim 1 to method described in any one of 46, wherein the level for measuring the UCH-L1 includes:

(1) antibody is captured, it is compound to form capture antibody-UCH-L1 antigen in conjunction with the epitope in UCH-L1 or UCH-L1 segment Object, and

(2) detect antibody, it includes detectable label and in conjunction on UCH-L1 not by it is described capture antibody combine epitope, with UCH-L1 antigen-detection antibody complex is formed,

(b) based on pass through it is described capture antibody-UCH-L1 antigen-detection antibody complex in the detectable label generate Signal measures the amount or concentration of UCH-L1 in the sample.

48. according to claim 1 to method described in any one of 47, wherein the sample is selected from the group being made up of: complete Blood sample, blood serum sample, spinal fluid samples and plasma sample.

49. according to claim 1 to method described in any one of 48, wherein in the subject by being shaken, led by body It causes the external mechanical force of closed or open head trauma or passivity impact that other power generate, one or many fall, is quick-fried The sample is obtained after head injury caused by fried or shock wave or other types of blunt forces wound.

50. according to claim 1 to method described in any one of 49, wherein taking in or being exposed to chemicals in the subject The sample is obtained after the combination of matter, toxin or chemical substance and toxin.

51. according to the method for claim 50, wherein the chemical substance or toxin be fire, mould, asbestos, insecticide, Insecticide, organic solvent, paint, glue, gas, organic metal, drug abuse or one or more combination.

52. according to claim 1 to method described in any one of 49, wherein the sample is from suffering from autoimmune disease, generation The subject for thanking disorder, brain tumor, anoxic, virus, meningitis, hydrocephalus or combinations thereof obtains.

53. wherein the method can be on anyone class subject according to claim 1 to method described in any one of 49 It carries out, without considering the factor in the group being made up of: the experiment of the clinical condition, human experimenter of human experimenter Room value, human experimenter as suffering from slight or moderate to the classification of severe TBI, the low or high UCH-L1 water of human experimenter Flat performance and human experimenter may suffer from the timing of any event of head injury.

54. according to claim 1 to method described in any one of 53, wherein the sample is biological sample.

55. according to claim 1 to method described in any one of 54, wherein the sample be whole blood sample, blood serum sample or Plasma sample.