CN104914254B - A kind of Platelet Detection - Google Patents
A kind of Platelet Detection Download PDFInfo
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- CN104914254B CN104914254B CN201410086000.8A CN201410086000A CN104914254B CN 104914254 B CN104914254 B CN 104914254B CN 201410086000 A CN201410086000 A CN 201410086000A CN 104914254 B CN104914254 B CN 104914254B
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
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Abstract
The present invention relates to blood testing technical field, particularly relate to a kind of Platelet Detection.This Platelet Detection carries out dissolving preparation by buffer after including mixing fibrinogen activator and platelet activating agent and forms mix reagent；In mix reagent, add antioxidation reagent form antioxidation mixing activation reagent；Antioxidation mixing is activated reagent put in specimen cup；Specimen cup is put in cillin bottle, and this cillin bottle is carried out freezing, evacuation and gland process；Being taken out from cillin bottle by specimen cup, be placed in thrombelastogram instrument, the whole blood sample after gathering adds in specimen cup；Start thrombelastogram instrument the whole blood sample being mixed with antioxidation mixing activation reagent in specimen cup is detected；Fibrinogenic intensity level is removed to obtain hematoblastic intensity level from Fibrinogen and hematoblastic combined strength bination value.The method is easy and simple to handle, testing result is not high by external interference and testing result accuracy.
Description
Technical field
The present invention relates to blood testing technical field, particularly relate to a kind of Platelet Detection.
Background technology
Thrombelastogram was invented by German Harter in 1948.The 1980's started to be widely used in
In clinical guidance art, blood transfusion achieves good result, has become current monitoring coagulation function
Important indicator.Thrombelastogram instrument can export blood clotting time R, blood clotting speed Angel and clot
The coagulation parameters such as intensity MA and the whole process of fibrinolytic.The physics of thrombelastogram monitoring blood clot
Characteristic is based on the principle that by a special static cylindrical cup filling blood with the angle of 4 ° 45 '
Rotate, rotate each time and continue 10 seconds.Hung by taenidium by one and be immersed in blood sample
Pin monitor the motion of blood sample.Cup and pin are sticked together by fibrin and platelet complex
After, cup rotates produced revolving force can be transferred to the pin in blood sample.Fibrin and platelet
The intensity of complex can affect the amplitude of needle movement, so that strong blood clot can make the motion of pin
Carry out with the synchronized movement of cup.Therefore, the motion amplitude of pin and the intensity of established blood clot
There is direct relation.When blood clot retraction or dissolving, pin releases with the connection of blood clot, cup
Motion is no longer transmitted to pin.The rotation of pin is converted into electronic signal by pickoff, this electricity
Subsignal can be monitored with computer.
At present, Fibrinogen and platelet function assay generally by thrombelastogram test into
Row.Wherein, thrombelastogram platelet function assay reagent be by common cup detectable,
Fibrin activator, platelet activating agent (such as arachidonic acid) form.Existing blood
The method of platelet Function detection includes: when Clinical detection, and Extemporaneous Fibrinogen activates
Both are also mixed to form mix reagent by agent and platelet activating agent；Mix reagent is put into XiLin
Lyophilized powder it is prepared as in Ping；As required quantitative mix reagent distilled water is redissolved；Multiple
Mix reagent after molten adds whole blood sample；By thrombelastogram instrument, whole blood sample is moved
State process of setting is monitored, thus quantitative scoring calculates the Fibrinogen of patient and hematoblastic
Combined strength bination；Fibrinogenic intensity is recorded, from Fibrinogen by thrombelastogram instrument
With hematoblastic combined strength bination is removed fibrinogenic intensity can obtain hematoblastic by force
Degree.
But, existing by thrombelastogram experiment carry out platelet function assay time, at sample
During detection can with weighing, calculate, redissolve, repeatedly multistep Reagent reconstitution and the examination such as sample-adding
Agent sample-adding process, operates extremely complex, and methods and results specificity is low before, and testing result is frequent
Being disturbed by external operation, its detection accuracy is poor simultaneously, and platelet activating agent is (such as flower
Raw tetraenoic acid) it is easy to oxidation, cause platelet activation to lose efficacy, there will be in Clinical detection
Fibrinogen false negative result, causes erroneous judgement to clinic, instructs the effect of patient medication
The best.
Therefore, for above not enough, the invention provides a kind of Platelet Detection.
Summary of the invention
(1) to solve the technical problem that
It is an object of the invention to provide a kind of Platelet Detection to solve the inspection of existing platelet
The operation that survey method exists is complicated, testing result is easily asked by external interference and detection accuracy difference
Topic.
(2) technical scheme
In order to solve above-mentioned technical problem, the invention provides a kind of Platelet Detection, its
Comprise the following steps:
S1, fibrinogen activator and platelet activating agent are mixed after carried out by buffer
Dissolve preparation and form mix reagent；
S2, in mix reagent add antioxidation reagent formed antioxidation mixing activate reagent；
S3, by antioxidation mixing activate reagent put in specimen cup；
S4, specimen cup is put in cillin bottle, and this cillin bottle is carried out freezing so that specimen cup
In antioxidation mixing activate reagent lyophilizing and stick to the cup of specimen cup at the bottom of；
S5, cillin bottle in step S4 is carried out evacuation and gland process, so that specimen cup is very
Altitude stores；
S6, specimen cup is taken out from cillin bottle, be placed in thrombelastogram instrument, after gathering
Whole blood sample add in specimen cup；
S7, startup thrombelastogram instrument activate reagent to being mixed with antioxidation mixing in specimen cup
Whole blood sample detects, quantitatively to draw Fibrinogen and hematoblastic combined strength bination value.
S8, remove from Fibrinogen and hematoblastic combined strength bination value fibrinogenic by force
Angle value is to obtain hematoblastic intensity level.
Wherein, in step S6, in whole blood sample being added specimen cup by sample loading gun and mix
Inhale, be sufficiently mixed with whole blood sample so that antioxidation mixing activates reagent.
Wherein, in step S1, described platelet activating agent is arachidonic acid, arachidonic acid
Concentration in mix reagent is 1～10mol/L.
Wherein, described buffer reagent includes trishydroxymethylaminomethane, phosphate buffer, second sulphur
One or more in acid and citric acid, the pH value of described mix reagent is 7.0～8.0.
Wherein, described fibrinogen activator includes thrombin, snake venom thrombin-like enzyme, Ba Qu
In enzyme and reptilase one or more.
Wherein, in step S1, described fibrinogen activator concentration in mix reagent is
0.1～10U/ml.
Wherein, in step S2, described antioxidation reagent include ascorbic acid, arabo-ascorbic acid,
One or more in vitamin E, thioctic acid and dithiothreitol, DTT, described antioxidation reagent is anti-
The concentration that oxidation mixing activates in reagent is 1～15mol/L.
Wherein, in step s3, put after antioxidation mixing activates and is mixed into solid-phase reagent in reagent
Enter in specimen cup.
Wherein, described solid-phase reagent includes sucrose, trehalose, lactose, maltose, glucose
With one or more in Polyethylene Glycol, solid-phase reagent mixes the volume activating reagent with antioxidation
Ratio range is 1%-10%.
(3) beneficial effect
The technique scheme of the present invention has the advantage that the platelets analysis that the present invention provides
In method, carried out by buffer after fibrinogen activator and platelet activating agent are mixed
Dissolve preparation and form mix reagent, be then mixed into the formation antioxidation mixing of antioxidation reagent and activate examination
After agent, then add fixating reagent wherein, prepared reagent is put in specimen cup, specimen cup
Put in cillin bottle, cillin bottle is carried out evacuation and gland processes, so that specimen cup is in vacuum
Environment stores；During Clinical detection, directly take out equipped with the cillin bottle in specimen cup, beat
Open cillin bottle taking-up specimen cup by thrombelastogram instrument, whole blood sample to be tested, can
Obtaining platelet test result easily, the method is easy and simple to handle, it is not necessary to weigh, calculate,
Redissolve, the repeatedly troublesome operation step such as sample-adding, testing result do not disturbed by external operation, antioxygen
Agent can effectively reduce the oxidation rate of platelet activating agent, makes platelet activating agent be in one
In inert environments, the sample in specimen cup is under vacuum environment when storage simultaneously, improves sample
The stability of reagent in product cup, and then the accuracy of platelets analysis result can be improved.
Accompanying drawing explanation
Fig. 1 is that in embodiment of the present invention Platelet Detection, the structure of specimen cup and cillin bottle is cutd open
View.
In figure, 1: cillin bottle；2: specimen cup；3: bowl cover；4: sealing-plug；5: dome；
6: easy-open lid.
Detailed description of the invention
Make to retouch the most in detail to the detailed description of the invention of the present invention with embodiment below in conjunction with the accompanying drawings
State.Following example are used for illustrating the present invention, but are not limited to the scope of the present invention.
In describing the invention, except as otherwise noted, " multiple " is meant that two kinds or two kinds
Above.Term " on ", D score, " interior ", the orientation of the instruction such as " outward " or position relationship be base
In orientation shown in the drawings or position relationship, it is for only for ease of the description present invention and simplifies description,
Rather than indicate or imply that the device of indication or element must have specific orientation, with specifically
Azimuth configuration and operation, be therefore not considered as limiting the invention.
Embodiment one
The Platelet Detection that the present invention provides comprises the following steps:
S1, fibrinogen activator and platelet activating agent are mixed after carried out by buffer
Dissolve preparation and form mix reagent；
Wherein, the pH value of mix reagent is 7.0-8.0；Fibrinogen activator include thrombin,
In snake venom thrombin-like enzyme, batroxobin and reptilase one or more, fibrinogen activator exists
Concentration in mix reagent is 0.1～10U/ml；Platelet activating agent is arachidonic acid, Semen arachidis hypogaeae
Tetraenoic acid concentration in mix reagent is 1～10mol/L；Buffer reagent includes trihydroxy methyl ammonia
One or more in methylmethane (Tris), phosphate buffer (PBS), ethyl sulfonic acid and citric acid.
S2, in mix reagent add antioxidation reagent formed antioxidation mixing activate reagent；
Wherein, antioxidation reagent includes ascorbic acid, arabo-ascorbic acid, vitamin E, thioctic acid
With one or more in dithiothreitol, DTT, described antioxidation reagent activates examination in antioxidation mixing
Concentration in agent is 1～15mol/L；Antioxidant can effectively reduce arachidonic oxidation speed
Rate, makes arachidonic acid be in an inert environments.
S3, by antioxidation mixing activate reagent put in specimen cup；Specimen cup includes glass body and cup
Lid, this specimen cup preferably can be placed on thrombelastogram instrument, is easy to the sample of experiment test
Cup.
S4, specimen cup is put in cillin bottle, and this cillin bottle is carried out freezing so that specimen cup
In antioxidation mixing activate reagent lyophilizing and stick to the cup of specimen cup at the bottom of；
Wherein, cillin bottle is sealed with sealing-plug, cillin bottle is placed on freeze dryer pallet,
Starting freeze dryer and carry out lyophilizing, freeze-drying time is 24 hours, wherein can put on freeze dryer simultaneously
Put multiple cillin bottle equipped with specimen cup.
S5, cillin bottle in step S4 is carried out evacuation and gland process, so that specimen cup is very
Altitude stores；
Wherein, vacuum is preferably 5～10Pa.So, it is ensured that freeze-dried reagent and external environment
Separate, it is to avoid reagent lost efficacy because of the oxygen in ingress of air and moisture, stabilized the property of reagent
Can, the beneficially long term storage of reagent.
S6, specimen cup is taken out from cillin bottle, be placed in thrombelastogram instrument, after gathering
Whole blood sample add in specimen cup；
S7, startup thrombelastogram instrument activate reagent to being mixed with antioxidation mixing in specimen cup
Whole blood sample detects, quantitatively to draw Fibrinogen and hematoblastic combined strength bination value.
Fibrinogen and hematoblastic combined strength bination value are by Fibrinogen and hematoblastic amplitude
Value embodies.
S8, according to existing conventional means obtain fibrinogenic intensity level, from fibrin
Former and hematoblastic combined strength bination value removes fibrinogenic intensity level hematoblastic to obtain
Intensity level.
When platelets analysis is tested, can directly fetch the cillin bottle that Vacuum Pressure in step S5 is built,
Open cillin bottle, take out specimen cup therein, specimen cup is placed on thrombelastogram instrument,
Then the whole blood of patient is acquired, the whole blood sample gathered is injected in specimen cup, carries out
Detection can obtain Fibrinogen and hematoblastic combined strength bination value, then removes by routine
Means obtain fibrinogenic intensity level, i.e. can obtain hematoblastic intensity level, it is achieved right
Hematoblastic detection.
In actual applications, by the present invention's after patient takes antiplatelet drug aspirin
Method detects hematoblastic function value, is then cut by the Fibrinogen function value of patient,
Can be obtained by the platelet function net value after patient's Aspirin, by not taking with patient
Medicine platelet net value contrast, can draw patient after the tablet has been ingested platelet suppression ratio, thus
Connect reflection patient's antiplatelet drug therapeutic effect of aspirin, instruct cardiovascular patient clinic to control
Treat and more after.
During Clinical detection, reagent is activated without adding, it is not necessary to enter by the method for the present invention
The preprocessing process of row blood preparation, can directly detect after blood specimen collection.Thrombelastogram instrument meeting
Automatically analyze the detection platelet amplitude of patient whole blood's sample, fibrin amplitude, thrombin shake
Width, can draw the antiplatelet drug suppression ratio of patient, quantitative analysis by particular analysis formula
Patient drug's action effect, quickly instruct cardio-cerebral vascular disease patient clinical diagnosis, treatment and more after,
Necessary drug effect monitoring foundation is provided for clinic.
Further, in step s 6, whole blood sample added in specimen cup also by sample loading gun
Carry out mixed suction, be sufficiently mixed with whole blood sample so that antioxidation mixing activates reagent.Wherein, mixed
The number of times inhaled is generally three times.
Preferably, in step s3, after antioxidation mixing activates and is mixed into solid-phase reagent in reagent
Put in specimen cup；
Wherein, described solid-phase reagent includes sucrose, trehalose, lactose, maltose, glucose
With one or more in Polyethylene Glycol (PEG), solid-phase reagent mixes activation examination with antioxidation
The volume ratio scope of agent is 1%-10%.Solid-phase reagent can fully be tied with specimen cup bottom even
Close, and this solid-phase reagent mix with blood after have good diffusion effect, specimen addition after
Quickly dissolve in blood, it is ensured that the stability of blood examination.
Embodiment two
As it is shown in figure 1, on the basis of embodiment one, embodiment two discloses a kind of platelet
The specimen cup 2 of detection method and cillin bottle 1.
Specifically, specimen cup 2 includes that glass body and bowl cover 3, bowl cover 3 main body are hollow circular cylinder,
Cylinder top end has circumferentially extending edge, and bowl cover 3 main body extend in glass chamber；Specimen cup 2
Being contained in cillin bottle 1, the bottleneck of cillin bottle 1 is provided with sealing-plug 4, the lower side pressure of sealing-plug 4
Being contained on the bowl cover 3 of specimen cup, cillin bottle 1 upper end is equipped with easy-open lid 6, easy-open lid 6 and sealing
Being provided with dome 5 between plug 4, the cavity in the bowl cover 3 of specimen cup is used for fixing metal needle,
Coordinate the use of thrombelastogram instrument；The lower end of sealing-plug 4 is fitted on the bowl cover 3 of specimen cup,
Prevent specimen cup 2 from moving during specimen cup 2 and cillin bottle 1 store and shift.
During use, first specimen cup 2 is positioned in cillin bottle 1；Secondly, by fibrin
Former activation reagent, platelet activation reagent and buffer are quantitatively adding the cavity in specimen cup 2
In, and the bowl cover 3 of specimen cup is contained in the cup of specimen cup；Then, by specimen cup 2 He
Cillin bottle 1 is positioned on freeze dryer and carries out lyophilizing；Finally, sealing-plug 4 is pressed under vacuum
It is packaged, then loads onto dome 5 and easy-open lid 6.
In sum, in the Platelet Detection that the present invention provides, Fibrinogen is activated
Carry out dissolving preparation by buffer after agent and arachidonic acid mixing and form mix reagent, then
After being mixed into antioxidation reagent formation antioxidation mixing activation reagent, then add fixating reagent wherein,
Putting in specimen cup by prepared reagent, specimen cup is put in cillin bottle, takes out cillin bottle
Vacuum and gland process, so that specimen cup stores in vacuum environment；During Clinical detection,
Directly take out equipped with the cillin bottle in specimen cup, open cillin bottle taking-up specimen cup and can pass through blood
Whole blood sample is tested by bolt elastic force figure instrument, obtains platelet test result, the party easily
Method is easy and simple to handle, it is not necessary to weighs, calculate, redissolve, the repeatedly troublesome operation step such as sample-adding,
Testing result is not disturbed by external operation, and antioxidant can effectively reduce arachidonic oxidation
Speed, makes arachidonic acid be in an inert environments, improves detection accuracy；Therein
Solid-phase reagent can fully be combined with specimen cup bottom even, and this solid-phase reagent mixes with blood
After have good diffusion effect, blood sample add after quickly dissolve in blood, it is ensured that
The stability of blood examination.
The above is only the preferred embodiment of the present invention, it is noted that lead for this technology
For the those of ordinary skill in territory, on the premise of without departing from the technology of the present invention principle, it is also possible to
Making some improvement and modification, these improve and modification also should be regarded as protection scope of the present invention.
Claims (2)
1. a Platelet Detection, it is characterised in that: it comprises the following steps:
S1, will fibrinogen activator and platelet activating agent mixing after by buffer carry out dissolve preparation formation mix reagent；
S2, in mix reagent add antioxidation reagent formed antioxidation mixing activate reagent；
S3, by antioxidation mixing activate reagent put in specimen cup；
S4, specimen cup is put in cillin bottle, and this cillin bottle is carried out freezing so that antioxidation mixing in specimen cup activate reagent lyophilizing and stick to specimen cup cup at the bottom of；
S5, cillin bottle in step S4 is carried out evacuation and gland process, so that specimen cup stores in vacuum environment；
S6, being taken out from cillin bottle by specimen cup, be placed in thrombelastogram instrument, the whole blood sample after gathering adds in specimen cup；
The whole blood sample being mixed with antioxidation mixing activation reagent in specimen cup is detected, quantitatively to draw Fibrinogen and hematoblastic combined strength bination value by S7, startup thrombelastogram instrument；
S8, from Fibrinogen and hematoblastic combined strength bination value, remove fibrinogenic intensity level to obtain hematoblastic intensity level；
In step S1, described platelet activating agent is arachidonic acid, and arachidonic acid concentration in mix reagent is 1～10mol/L；
Described buffer reagent includes that one or more in trishydroxymethylaminomethane, phosphate buffer, ethyl sulfonic acid and citric acid, the pH value of described mix reagent are 7.0～8.0；
Described fibrinogen activator includes one or more in thrombin, snake venom thrombin-like enzyme, batroxobin and reptilase；
In step S1, described fibrinogen activator concentration in mix reagent is 0.1～10U/ml；
In step S2, described antioxidation reagent includes one or more in ascorbic acid, arabo-ascorbic acid, vitamin E, thioctic acid and dithiothreitol, DTT, and the concentration that described antioxidation reagent activates in reagent in antioxidation mixing is 1～15mol/L；
In step s3, put in specimen cup after antioxidation mixing activates and is mixed into solid-phase reagent in reagent；
Described solid-phase reagent includes one or more in sucrose, trehalose, lactose, maltose, glucose and Polyethylene Glycol, and it is 1%-10% that solid-phase reagent mixes the volume ratio scope of activation reagent with antioxidation.
Platelet Detection the most according to claim 1, it is characterised in that: in step S6, in whole blood sample being added specimen cup by sample loading gun and carry out mixed suction, it is sufficiently mixed so that antioxidation mixing activates reagent with whole blood sample.
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109884326B (en) * | 2019-03-27 | 2022-08-19 | 深圳优迪生物技术有限公司 | Platelet aggregation function detection kit |
| CN110514823B (en) * | 2019-08-28 | 2023-12-05 | 深圳麦科田生物医疗技术股份有限公司 | Platelet inhibition rate detection kit, preparation method and detection method thereof |
| CN113156144A (en) * | 2021-01-29 | 2021-07-23 | 郑州普湾医疗技术有限公司 | Arachidonic acid cup detection reagent with platelet aggregation function and preparation method thereof |
| CN112730769A (en) * | 2021-01-29 | 2021-04-30 | 郑州普湾医疗技术有限公司 | Platelet aggregation functional adenosine diphosphate cup detection reagent and preparation method thereof |
| CN112924701A (en) * | 2021-01-29 | 2021-06-08 | 郑州普湾医疗技术有限公司 | Batroxobin cup detection reagent with platelet aggregation function and preparation method thereof |
| CN113848332B (en) * | 2021-09-17 | 2024-04-19 | 广州徕西姆医学诊断技术有限公司 | Thrombus elastography detection reagent and preparation method and application thereof |
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| CN102980993A (en) * | 2012-11-06 | 2013-03-20 | 北京乐普医疗科技有限责任公司 | Platelet aggregation function detection kit and detection method |
| CN203191377U (en) * | 2013-04-02 | 2013-09-11 | 北京乐普医疗科技有限责任公司 | Platelet aggregation function detection device |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102980993A (en) * | 2012-11-06 | 2013-03-20 | 北京乐普医疗科技有限责任公司 | Platelet aggregation function detection kit and detection method |
| CN203191377U (en) * | 2013-04-02 | 2013-09-11 | 北京乐普医疗科技有限责任公司 | Platelet aggregation function detection device |
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Address after: 102200, Beijing Changping District science and Technology Park, super Road, No. 7-1, building 37 Patentee after: Beijing Lepu Diagnostic Technology Co., Ltd Address before: 102200, Beijing Changping District science and Technology Park, super Road, No. 7-1, building 37 Patentee before: BEIJING LEPU MEDICAL TECHNOLOGY Co.,Ltd. |