CN109212225A - A kind of phosphorylated Tau protein detection reagent item and its application - Google Patents
A kind of phosphorylated Tau protein detection reagent item and its application Download PDFInfo
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Abstract
The present invention provides a kind of phosphorylated Tau protein detection reagent item and its application, the reagent strip (1) includes the liquid storage cavity layer set gradually from top to bottom, flow channel layer and reaction cavity layer；The liquid storage cavity layer is provided at least one liquid storage chamber (4)；The reaction cavity layer is provided at least one reaction chamber (2)；The flow channel layer is provided with microchannel (8), for being connected to liquid storage chamber (4) and reaction chamber (2).Reagent strip of the invention is based on microflow control technique, it is integrated to will test reagent, avoid reagent contamination, reduce transportation cost, the reagent strip is loaded on detecting instrument, automation Molecular Detection is realized, experimental error caused by open system pollution that may be present and manual operation is eliminated, there is important application in phosphorylated Tau protein early detection.
Description
Technical field
The invention belongs to Protein Detection field, it is related to a kind of phosphorylated Tau protein detection reagent item and its application, especially relates to
And a kind of phosphorylated Tau protein detection reagent item and its application based on magnetic particle and chemiluminescence.
Background technique
According to statistics, there is nearly 30,000,000 alzheimer's disease (Alzheimer disease, AD) patient in the whole world.The sea A Erzi
Silent disease makes a definite diagnosis rear mean survival time (MST) only 10 years, still falls within what medical field can not cure at present to the harm of the elderly and its serious
Pernicious old age mental disease.For 60 years old or more crowd, with advancing age, the disease incidence of alzheimer's disease was sharply risen
It is high.In China, the alzheimer's disease disease incidence of over-65s crowd is 7%, and the disease incidence of 85 years old or more crowd is 25%, and
The disease incidence of 95 years old or more crowds is then up to 60%.Early stage alzheimer's disease non-evident sympton, it is difficult to diagnose, easily be missed
It examines.
Alzheimer's disease research association thinks that the lesion growth of alzheimer's disease is slow, early diagnosis, early intervention
The generation that lesion can significantly be delayed, it is most important for reducing disease incidence, be conducive to the normal life for maintaining patient.Meanwhile
Early diagnosis helps to distinguish Alzheimer's and the doubtful patient of alzheimer's disease, improves diagnostic accuracy.
Microtubule system is the main component of nerve cell skeleton, is played a significant role on maintaining cell function.Tau albumen
It is a kind of microtubule associated protein, in conjunction with micro-pipe and the stability of micro-pipe can be kept.Under normal circumstances, each Tau albumen contains
2-3 phosphorylation site, and the content of the phosphorylated Tau protein (P-Tau) of Alzheimer's intracerebral is higher than normal person
3-4 times, excessive P-Tau assembles to form neurofibrillary tangles (NFT), causes the serious damage of nerve cell, influences normal
Brain function.It is respectively normal phosphorylation Tau, solvable in Alzheimer's intracerebral there are three kinds of phosphorylated Tau proteins
Sexual abnormality phosphorylated Tau protein and the abnormal Phosphorylation Tau for being gathered into PHF (conjugate spirals silk), wherein title phosphoric acid has been reported
Changing Tau has 45 phosphorylation sites, and including Thr181, Thr231, Ser396, Ser404 etc., the detection of phosphorylation site indicates
The raising of alzheimer's disease illness possibility, therefore, detection P-Tau content facilitate early screening alzheimer's disease trouble
Person.
101307108 A of CN discloses anti-phosphorylated Tau protein antibody and its in AD disease abnormal Phosphorylation Tau albumen water
Purposes in flat measurement prepares each group brain tissue paraffin section by establishing AD disease rat model；Using resisting each site phosphorus respectively
The antibody of acidification Tau albumen does primary antibody；Phosphorylated Tau protein in each group rat cerebral tissue nerve cell is detected, is as a result detected
Each site phosphorylation Tau albumen.However the early stage that this method is not particularly suited for phosphorylated Tau protein is quickly detected.
Therefore it provides a kind of high sensitivity, high specificity, the phosphorylation Tau egg strong with automatic detection system compatibility
White detection reagent item is of great significance in alzheimer's disease research field.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of phosphorylated Tau protein detection reagent item and its application, described
Reagent strip is based on microflow control technique and magnetic microparticle chemiluminescence technology, and high specificity, high sensitivity, accuracy are good, phosphorylation Tau
Protein assay reagent is loaded on reagent strip, realizes the integrated of reagent strip, is matched with equipment is tested and analyzed, real
Automation, highly sensitive, the quick detection for having showed phosphorylated Tau protein, are that the early diagnosis field of alzheimer's disease has
Significance.
To achieve this purpose, the present invention adopts the following technical scheme:
In a first aspect, the reagent strip 1 includes the liquid storage set gradually from top to bottom the present invention provides a kind of reagent strip
Cavity layer, flow channel layer and reaction cavity layer；
The liquid storage cavity layer is provided at least one liquid storage chamber 4；
The reaction cavity layer is provided at least one reaction chamber 2；
The flow channel layer is provided with microchannel 8, for being connected to liquid storage chamber 4 and reaction chamber 2；
Detection reagent is housed in the liquid storage chamber 4；
The detection reagent includes that Tau albumen capture antibody coupling magnetic particle, cleaning solution, phosphorylated Tau protein detection are anti-
In body or reaction solution any one or at least two combination.
Reagent strip of the invention not only increases detection efficiency using microchannel connection liquid storage chamber and reaction chamber, and
The consumption for reducing sample and reagent is conducive to be loaded into and realizes that the automation of molecule is controllably examined on automatic detection device
It surveys.
Preferably, the reagent strip 1 further includes at least one sealing plug 3, is loaded into liquid storage chamber 4.
In the present invention, sealing plug is placed in liquid storage chamber, the leakproofness of liquid storage chamber is not only ensure that, avoids detection reagent
It is contaminated in storage and transportational process, and is cooperated by sealing plug and power pressurizing unit, by the inspection in liquid storage chamber
Test agent indentation microchannel enters reaction chamber, realizes the detection function of reagent strip.
Preferably, at least one buckle 6 is provided on the outer surface of the sealing plug 3, on the inner surface of the liquid storage chamber 4
At least two card slots 7 are provided with, the position of sealing plug 3 is limited by connecting with buckle 6.
In the present invention, by buckling the connection with card slot, position of the sealing plug in liquid storage chamber is defined, when on sealing plug
Buckle be connected with the first card slot in liquid storage chamber, sealing plug is in liquid storage position, and detection reagent does not flow out, when on sealing plug
Buckle is connected with the second card slot in liquid storage chamber, and sealing plug is in liquid release position, and sealing plug, which squeezes liquid storing bag, makes liquid storing bag
Rupture, detection reagent outflow.
Preferably, the length of the reaction cavity layer is not less than the length of the liquid storage cavity layer.
In the present invention, sample to be tested is added as well relative to the part that liquid storage cavity layer is protruded in reaction cavity layer.
Preferably, the reaction cavity layer is provided with partition 9, and the height of the partition 9 is less than the height of the reaction cavity layer.
Reaction chamber interlayer is divided into different reaction chambers, it is therefore prevented that phase by the way that partition is arranged in reaction cavity layer by the present invention
Cross contamination occurs for adjacent reagent, increases the accuracy of reagent strip；In addition, the height of partition is less than the height of reaction cavity layer, make
On partition top, there are appropriate spaces, moved between differential responses chamber for magnetic particle.
Preferably, the upper surface of the partition 9 includes any one in plane, inclined-plane or curved surface, preferably inclined-plane.
Preferably, the bottom surface of the reagent strip 1 is provided with locating slot 5, for the reagent strip 1 to be fixed on detecting instrument
On.
Preferably, the volume of the liquid storage chamber 4 be 10-500 μ L, such as can be 10 μ L, 50 μ L, 100 μ L, 150 μ L,
200 μ L, 250 μ L, 300 μ L, 350 μ L, 400 μ L, 450 μ L or 500 μ L.
Preferably, the volume of the reaction chamber 2 be 10-500 μ L, such as can be 10 μ L, 50 μ L, 100 μ L, 150 μ L,
200 μ L, 250 μ L, 300 μ L, 350 μ L, 400 μ L, 450 μ L or 500 μ L.
Preferably, the reagent strip 1 is molded to be formed using high molecular material.
Preferably, the high molecular material includes acrylonitrile-butadiene-styrene copolymer, polymethyl methacrylate
In polyethylene any one or at least two combination, preferably polymethyl methacrylate.
Preferably, detection reagent is housed, the detection reagent is loaded in sealing liquid storing bag in the liquid storage chamber 4.
Preferably, the sealing liquid storing bag is using any one in plastic film, aluminium foil or aluminium plastic composite material or extremely
Few two kinds of combination is prepared, and is preferably prepared using aluminium plastic composite material.
Sealing liquid storing bag of the invention is using the sealing means sealing being easy to tear, under the pressure effect of sealing plug, storage
The rupture of liquid bag, realizes the release of detection reagent.
Preferably, the detection reagent includes Tau albumen capture antibody coupling magnetic particle, cleaning solution, phosphorylated Tau protein
Detect in antibody or reaction solution any one or at least two combination, preferably Tau albumen capture antibody coupling magnetic particle,
The combination of cleaning solution, phosphorylated Tau protein detection antibody and reaction solution.
Preferably, the partial size of Tau albumen capture antibody coupling magnetic particle is 0.1-10 μm, for example, can be 0.1 μm,
0.5 μm, 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm or 10 μm.
Magnetic particle surface modification of the invention has Tau protein antibodies, and the antibody is to non-phosphorylating Tau albumen and phosphorylation
Tau albumen all has binding function, after sample and magnetic particle are incubated for jointly, non-phosphorylating Tau albumen and phosphoric acid in sample
Change Tau protein binding on magnetic particle surface, antibody is further detected using phosphorylated Tau protein, can be detected to obtain phosphorylation
Tau albumen.
Preferably, the detecting instrument includes power pressurizing unit, magnetic particle mobile controller or signal supervisory instrument
In any one or at least two combination, preferably power pressurizing unit, magnetic particle mobile controller and signal detection
The combination of device.
In the present invention, the course of work of the reagent strip is as follows:
(1) by Tau albumen capture antibody coupling magnetic particle solution, cleaning solution, phosphorylated Tau protein detection antibody-solutions and
Reaction solution is respectively charged into liquid storing bag sealing, is respectively placed in corresponding liquid storage chamber 4；
(2) when reagent strip 1 stores and transports, sealing plug 3 is inserted into liquid storage chamber 4, makes buckle 6 and storage on sealing plug 3
The first card slot connection in sap cavity 4, sealing plug 3 are in liquid storage position；
(3) in an experiment, reagent strip 1 is put into first and is tested and analyzed in instrument, the power pressurizing unit of instrument squeezes envelope
Mouth plug 3, connect the buckle 6 on sealing plug 3 with the second card slot in liquid storage chamber, and liquid storing bag rupture, detection reagent passes through miniflow
Channel 8 flows into reaction chamber 2；
(4) then to sample to be tested is added in the first reaction chamber of reagent strip 1, it is micro- that antibody coupling magnetic is captured with Tau albumen
Grain is incubated for；
(5) the magnetic particle mobile controller of instrument by the first reaction chamber magnetic particle enrichment after, by reaction chamber it
Between 9 top of partition space, magnetic particle is transferred to the second reaction chamber, carries out magnetic particle cleaning；
(6) same steps are used, magnetic particle is transferred to adjacent reaction chamber, the signal detection dress of ultimate analysis instrument respectively
Acquisition signal is set, output analysis result after software processing.
Second aspect, the present invention provides a kind of reagent strip as described in relation to the first aspect detection phosphorylated Tau protein and/
Or phosphorylated Tau protein passes through replacing, missing or adding for individual amino acid residue and obtains living with phosphorylated Tau protein
Application in the amino acid sequence of property.
Compared with prior art, the invention has the following beneficial effects:
(1) reagent strip of the invention utilizes microflow control technique, is connected to liquid storage chamber and reaction chamber using microchannel, not only mentions
High detection efficiency, and reduce the consumption of sample and reagent, be conducive to be loaded on automatic detection device and realize phosphorus
The automation of acidification Tau albumen controllably detects；
(2) detection reagent in reagent strip of the invention is integrated in the reagent strip, using sealing plug to liquid storing bag
The detection reagent being sealed in liquid storing bag is discharged into reaction chamber by squeezing action, avoids reagent contamination, reduce transport at
This；
(3) reagent strip of the invention is loaded on detecting instrument, other than sample to be tested is added, all experimental implementations,
The transfer of release, magnetic particle including detection reagent and the detection of signal, are automatically performed by analysis instrument, easy to operate easy
Row eliminates experimental error caused by open system pollution that may be present and manual operation, examines in phosphorylated Tau protein early stage
There is important application in survey.
Detailed description of the invention
Fig. 1 is the southeast isometric view of reagent strip of the present invention, wherein 1- reagent strip, 2- reaction chamber, 3- sealing plug, 4-
Liquid storage chamber, 5- locating slot；
Fig. 2 is the right side view of reagent strip of the present invention, wherein 3- sealing plug, 5- locating slot；
Fig. 3 is the isometric schematic cross-sectional view in the local southeast of reagent strip of the present invention, wherein 6- buckle, 7- card slot, 8- miniflow are logical
Road, 9- partition；
Fig. 4 is the isometric schematic cross-sectional view in the whole southeast of reagent strip of the present invention, wherein 10- magnetic particle liquid storage chamber, 11- the
One cleaning solution liquid storage chamber, 12- the second cleaning solution liquid storage chamber, 13- detect antibody stock chamber, 14- third cleaning solution liquid storage chamber, 15-
4th cleaning solution liquid storage chamber, 16- chemiluminescence reaction liquid liquid storage chamber, 17- sample incubation chamber, 18- first clean chamber, 19- second
Chamber is cleaned, 20- detects antibody incubation chamber, and 21- third cleans chamber, and 22- the 4th cleans chamber, 23- chemiluminescence detection chamber；
Fig. 5 is the standard curve using reagent strip of the present invention detection phosphorylated Tau protein concentration；
Fig. 6 is the chemiluminescence result using reagent strip of the present invention detection sample.
Specific embodiment
The technological means and its effect taken for the present invention is further explained, with reference to embodiments with attached drawing to this hair
It is bright to be further described.It is understood that the specific embodiments described herein are used only for explaining the present invention, rather than
Limitation of the invention.
In the examples where no specific technique or condition is specified, described technology or conditions according to the literature in the art,
Or it is carried out according to product description.Reagents or instruments used without specified manufacturer, be can be by regular channel commercially available from
The conventional products of acquisition.
The assembling of 1 reagent strip of embodiment
The southeast isometric view of the reagent strip of the present embodiment as shown in Figure 1, right side view as shown in Fig. 2, part east
The isometric schematic cross-sectional view in south is as shown in figure 3, the isometric schematic cross-sectional view in the whole southeast is as shown in Figure 4.
The reagent strip 1 includes the liquid storage cavity layer set gradually from top to bottom, flow channel layer and reaction cavity layer；
The liquid storage cavity layer is provided with seven liquid storage chambers 4, respectively magnetic particle liquid storage chamber 10, the first cleaning solution liquid storage chamber
11, the second cleaning solution liquid storage chamber 12, detection antibody stock chamber 13, third cleaning solution liquid storage chamber 14, the 4th cleaning solution liquid storage chamber 15
With chemiluminescence reaction liquid liquid storage chamber 16；
The reaction cavity layer is provided with seven reaction chambers 2, and it is clear to clean chamber 18, second for respectively sample incubation chamber 17, first
It washes chamber 19, detection antibody incubation chamber 20, third cleaning chamber the 21, the 4th and cleans chamber 22 and chemiluminescence detection chamber 23；
The flow channel layer is provided with seven microchannels 8, for being connected to liquid storage chamber and corresponding reaction chamber；
The length of the reaction cavity layer is greater than the length of the liquid storage cavity layer；
The reagent strip 1 further includes seven sealing plugs, is loaded into corresponding liquid storage chamber；
A buckle 6 is provided on the outer surface of the sealing plug 3, on the inner surface of the liquid storage chamber 4 there are two settings
Highly different card slots 7 limits the position of sealing plug 3 by connecting with buckle 6；
The reaction cavity layer is provided with partition 9, and reaction chamber interlayer is divided into reaction chamber 17,18,19,20,21,22 and 23,
The height of the partition 9 is less than the height of the reaction cavity layer；
The upper surface of the partition 9 is inclined-plane；
The bottom surface of the reagent strip 1 is provided with locating slot 5, for the reagent strip 1 to be fixed on detecting instrument；
The volume of the liquid storage chamber 4 is 10-500 μ L；
Preferably, the volume of the reaction chamber 2 is 10-500 μ L.
The course of work of reagent strip of the invention is as follows:
(1) Tau albumen is captured into antibody coupling magnetic particle solution, the first cleaning solution, the second cleaning solution, phosphorylation Tau egg
White detection antibody-solutions, third cleaning solution, the 4th cleaning solution and chemiluminescence reaction liquid are respectively charged into liquid storing bag sealing, put respectively
It is placed in magnetic particle liquid storage chamber 10, the first cleaning solution liquid storage chamber 11, the second cleaning solution liquid storage chamber 12, detection antibody stock chamber 13, the
In three cleaning solution liquid storage chambers 14, the 4th cleaning solution liquid storage chamber 15 and chemiluminescence reaction liquid liquid storage chamber 16；
(2) when reagent strip 1 stores and transports, sealing plug is inserted into liquid storage chamber, makes buckle 6 and liquid storage chamber on sealing plug
Interior the first card slot connection, sealing plug are in liquid storage position；
(3) in an experiment, reagent strip 1 is put into first and is tested and analyzed in instrument, the power pressurizing unit of instrument squeezes envelope
Mouth plug, ruptures liquid storing bag, detection reagent flows into reaction chamber by microchannel；
(4) it then to sample to be tested is added in the sample incubation chamber 17 of reagent strip, is incubated for magnetic particle；
(5) the magnetic particle mobile controller of instrument is incubated after the magnetic particle enrichment in sample incubation chamber 17 by sample
Tau albumen capture antibody coupling magnetic particle is transferred to the by the space for educating the partition top between chamber 17 and the first cleaning chamber 18
One cleaning chamber 18, carries out magnetic particle cleaning；
(6) same steps are used, Tau albumen capture antibody coupling magnetic particle is transferred to the second cleaning chamber 19, inspection respectively
Survey antibody incubation chamber 20, third cleaning chamber the 21, the 4th cleans chamber 22 and chemiluminescence detection chamber 23, the signal of ultimate analysis instrument
Detection device acquires the chemiluminescence signal of chemiluminescence detection chamber 23, output analysis result after software processing.
The detection of 2 phosphorylated Tau protein standard items of embodiment
The reagent strip of the present embodiment is formed using polymethyl methacrylate (PMMA) injection molding, and liquid storing bag is using plastic-aluminum combined
Material is prepared.
Respectively by 50 μ L Tau protein antibodies be coupled magnetic particle solution, 100 the first cleaning solutions of μ L, 100 the second cleaning solutions of μ L,
The anti-phosphorylated Tau protein antibody of 100 μ L alkali phosphatase enzyme marks, 100 μ L third cleaning solutions, the 4th cleaning solution of 100 μ L and 100 μ L
After alkaline phosphatase substrate AMPPD reaction solution injects liquid storing bag, is sealed using welding technique, be respectively placed in magnetic particle liquid storage chamber
10, the first cleaning solution liquid storage chamber 11, the second cleaning solution liquid storage chamber 12, detection antibody stock chamber 13, third cleaning solution liquid storage chamber 14,
In 4th cleaning solution liquid storage chamber 15 and chemiluminescence reaction liquid liquid storage chamber 16, the reagent strip is obtained.
The preparation of phosphorylated Tau protein standard items includes carrying the artificial synthesized gene constructed recombination of phosphorylated Tau protein
Body imports host strain, and Protein expression and purification is carried out after screening and is obtained, and obtains phosphorylation Tau egg using later period phosphorylation modification
It is white, phosphorylated Tau protein standard items using TBS or PBS buffer solution dilution mother liquor obtain, concentration be respectively 320pg/mL,
160pg/mL, 80pg/mL, 40pg/mL, 20pg/mL, 10pg/mL, 5pg/mL and 0pg/mL.
In the present embodiment, the sealed membrane of 8 reagent strips is torn off for the detection of primary achievable 8 reagent strips of detecting instrument,
The 100 μ L of phosphorylated Tau protein standard items of various concentration is added into magnetic particle liquid storage chamber 10, is put into detecting instrument starting inspection
It surveys, the reagent of liquid storage chamber is added in corresponding reaction chamber by power pressurizing unit；
It is 37 DEG C that the temperature control console of instrument, which keeps temperature locating for reagent strip, at this time phosphorylated Tau protein standard items with repair
0.1 μm of magnetic particle for being decorated with capture antibody reacts 10-15min at a suitable temperature；
Then, start magnetic particle transfer device, the magnetic particle absorption of instrument is with transfer device by sample by sample incubation chamber
Respectively by the first cleaning chamber and the second cleaning chamber, magnetic particle cleaning is carried out；
The magnetic particle cleaned is transferred to detection antibody incubation chamber, capture has the magnetic particle and alkali of phosphorylated Tau protein
Acid phosphatase marks anti-phosphorylated Tau protein antibody mixing, 37 DEG C of reaction 10-15min；
Start magnetic particle transfer device, sample is incubated for chamber by detection and led to respectively by the magnetic particle absorption of instrument and transfer device
Third cleaning chamber and the 4th cleaning chamber are crossed, magnetic particle cleaning is carried out；
The magnetic particle cleaned is transferred to chemiluminescence detection chamber, carries out chemiluminescence detection.
The values of chemiluminescence of phosphorylated Tau protein standard items is as shown in table 1 and Fig. 5, equation of linear regression y=
1.7099x+258.93 relative coefficient R2=0.9985, have between phosphorylated Tau protein concentration and values of chemiluminescence good
Good linear relationship, illustrates the reliability and accuracy of reagent strip and instrument.
Table 1
The detection of 3 phosphorylated Tau protein clinical sample of embodiment
The reagent strip of the present embodiment is formed using acrylonitrile-butadiene-styrene copolymer (ABS) injection molding, and liquid storing bag is adopted
It is prepared with aluminium plastic composite material.
Respectively by 50 μ L Tau protein antibodies be coupled magnetic particle solution, 100 the first cleaning solutions of μ L, 100 the second cleaning solutions of μ L,
The anti-phosphorylated Tau protein antibody of 100 μ L alkali phosphatase enzyme marks, 100 μ L third cleaning solutions, the 4th cleaning solution of 100 μ L and 100 μ L
After alkaline phosphatase substrate AMPPD reaction solution injects liquid storing bag, is sealed using welding technique, be respectively placed in magnetic particle liquid storage chamber
10, the first cleaning solution liquid storage chamber 11, the second cleaning solution liquid storage chamber 12, detection antibody stock chamber 13, third cleaning solution liquid storage chamber 14,
In 4th cleaning solution liquid storage chamber 15 and chemiluminescence reaction liquid liquid storage chamber 16, the reagent strip is obtained.
3000rpm/min at whole blood sample each 16,4 DEG C of Alzheimer's and normal person is chosen to be centrifuged
10min collects blood plasma or serum, -80 DEG C of preservations；
Sample is diluted 5 times using PBS or TBS buffer, 50 μ L is drawn and reagent strip is added, be put into instrument and examined
It surveys, as a result as shown in table 2 and Fig. 6, it can be seen that the detection that the detection average value (295.69) of patient is significantly higher than normal person is flat
Mean value (188.91), difference 56.52%, with the intracorporal phosphorylated Tau protein total amount of Alzheimer's reported in the literature
The result for being significantly higher than normal person is consistent, and illustrates that the testing result of the present embodiment reagent strip has referential.
2 sample typesetting of table and corresponding detection luminous value
In conclusion reagent strip of the invention utilizes microflow control technique, liquid storage chamber and reaction chamber are connected to using microchannel,
Detection efficiency is not only increased, and reduces the consumption of sample and reagent, is conducive to be loaded on automatic detection device
Realize that the automation of molecule controllably detects；Detection reagent is integrated in reagent strip, using sealing plug to the squeezing action of liquid storing bag
The detection reagent being sealed in liquid storing bag is discharged into reaction chamber, reagent contamination is avoided, reduces transportation cost；Reagent strip
It is loaded on detecting instrument, other than sample to be tested is added, all experimental implementations, release, magnetic particle including detection reagent
Transfer and signal detection, be automatically performed by analysis instrument, it is easy to operation, it is that may be present to eliminate open system
Experimental error caused by pollution and manual operation；Phosphorylated Tau protein detection, the inspection of patient are carried out using reagent strip of the invention
Survey average value (295.69) and be significantly higher than the detection average value (188.91) of normal person, difference 56.52%, with Ah. reported in the literature
The result that the phosphorylated Tau protein total amount of the silent disease patient's body in Wurz sea is significantly higher than normal person is consistent, and testing result has
Referential.
The Applicant declares that the present invention is explained by the above embodiments method detailed of the invention, but the present invention not office
Be limited to above-mentioned method detailed, that is, do not mean that the invention must rely on the above detailed methods to implement.Technical field
Technical staff it will be clearly understood that any improvement in the present invention, equivalence replacement and auxiliary element to each raw material of product of the present invention
Addition, selection of concrete mode etc., all of which fall within the scope of protection and disclosure of the present invention.
Claims (10)
1. a kind of reagent strip, which is characterized in that the reagent strip (1) includes the liquid storage cavity layer set gradually from top to bottom, runner
Layer and reaction cavity layer；
The liquid storage cavity layer is provided at least one liquid storage chamber (4)；
The reaction cavity layer is provided at least one reaction chamber (2)；
The flow channel layer is provided with microchannel (8), for being connected to liquid storage chamber (4) and reaction chamber (2)；
Detection reagent is housed in the liquid storage chamber (4)；
The detection reagent include Tau albumen capture antibody coupling magnetic particle, cleaning solution, phosphorylated Tau protein detection antibody or
In reaction solution any one or at least two combination.
2. reagent strip according to claim 1, which is characterized in that the reagent strip (1) further includes at least one sealing plug
(3), it is loaded into liquid storage chamber (4)；
Preferably, at least one buckle (6), the inner surface of the liquid storage chamber (4) are provided on the outer surface of the sealing plug (3)
On be provided at least two card slots (7), by connect with buckle (6) limit sealing plug (3) position；
Preferably, the length of the reaction cavity layer is not less than the length of the liquid storage cavity layer.
3. reagent strip according to claim 1 or 2, which is characterized in that the reaction cavity layer is provided with partition (9), described
The height of partition (9) is less than the height of the reaction cavity layer；
Preferably, the upper surface of the partition (9) includes any one in plane, inclined-plane or curved surface, preferably inclined-plane.
4. reagent strip according to claim 1-3, which is characterized in that the bottom surface of the reagent strip (1) is provided with
Locating slot (5), for the reagent strip (1) to be fixed on detecting instrument.
5. reagent strip according to claim 1-4, which is characterized in that the volume of the liquid storage chamber (4) is 10-
500μL；
Preferably, the volume of the reaction chamber (2) is 10-500 μ L.
6. reagent strip according to claim 1-5, which is characterized in that the reagent strip (1) uses macromolecule material
Material injection molding is formed；
Preferably, the high molecular material includes acrylonitrile-butadiene-styrene copolymer, polymethyl methacrylate or poly-
In ethylene any one or at least two combination, preferably polymethyl methacrylate.
7. reagent strip according to claim 1-6, which is characterized in that the detection reagent is loaded on sealing liquid storing bag
In；
Preferably, the sealing liquid storing bag uses any one in plastic film, aluminium foil or aluminium plastic composite material or at least two
The combination of kind is prepared, and is preferably prepared using aluminium plastic composite material.
8. reagent strip according to claim 1-7, which is characterized in that the detection reagent includes that Tau albumen is caught
Obtain the combination of antibody coupling magnetic particle, cleaning solution, phosphorylated Tau protein detection antibody and reaction solution；
Preferably, the partial size of the Tau albumen capture antibody coupling magnetic particle is 0.1-10 μm.
9. reagent strip according to claim 1-8, which is characterized in that the detecting instrument includes that power squeezes dress
Set, in magnetic particle mobile controller or signal supervisory instrument any one or at least two combination, preferably power squeeze
The combination of pressure device, magnetic particle mobile controller and signal supervisory instrument.
10. a kind of if the described in any item reagent strips of claim 1-9 are in detection phosphorylated Tau protein and/or phosphorylation Tau egg
It is white obtained by replacing, missing or adding for individual amino acid residue have the active amino acid sequence of phosphorylated Tau protein
Application in column.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110514827A (en) * | 2019-09-26 | 2019-11-29 | 天津华科泰生物技术有限公司 | A kind of immunochromatographydetection detection card and preparation method thereof of the total Tau albumen of quick detection |
| CN112162101A (en) * | 2020-10-13 | 2021-01-01 | 南京立顶医疗科技有限公司 | Kit for detecting biomarkers of Alzheimer's disease and detection method thereof |
| CN112505322A (en) * | 2020-10-16 | 2021-03-16 | 威海纽普生物技术有限公司 | Alzheimer disease marker p-Tau217 detection kit and manufacturing method thereof |
| CN113533746A (en) * | 2021-07-22 | 2021-10-22 | 深圳市天大生物医疗器械有限公司 | P-Tau protein chemiluminescence detection kit and preparation method thereof |
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| CN112162101A (en) * | 2020-10-13 | 2021-01-01 | 南京立顶医疗科技有限公司 | Kit for detecting biomarkers of Alzheimer's disease and detection method thereof |
| CN112505322A (en) * | 2020-10-16 | 2021-03-16 | 威海纽普生物技术有限公司 | Alzheimer disease marker p-Tau217 detection kit and manufacturing method thereof |
| CN113533746A (en) * | 2021-07-22 | 2021-10-22 | 深圳市天大生物医疗器械有限公司 | P-Tau protein chemiluminescence detection kit and preparation method thereof |
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