Kit for determining TAT content and preparation method thereof

A kit and reagent technology, applied in the field of immunoassay, can solve the problems of reagent stability interference, reagent freezing and thawing, reagent system instability, etc., and achieve the effects of good reagent stability, high detection sensitivity and broad application prospects

Active Publication Date: 2022-07-01
SHENZHEN DYMIND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Moreover, the TAT reagent will be affected by many factors during the storage process. For example, after freezing and thawing, the performance of the reagent will be destroyed and the reagent system will be unstable, which will affect the test sensitivity and the subsequent long-term stability of the reagent.
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Method used

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  • Kit for determining TAT content and preparation method thereof
  • Kit for determining TAT content and preparation method thereof
  • Kit for determining TAT content and preparation method thereof

Examples

Experimental program
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Effect test

Example Embodiment

[0036] The preparation method of the above-mentioned test kit comprises the following steps:

[0037] S1. Magnetically separate the carboxyl group-modified particle solution under the action of a magnetic field, remove the supernatant, wash with an activation buffer, then resuspend the washed magnetic particles with an activation buffer and add a first activator to activate to obtain activated magnetic particles .

[0038] S2. Take the activated magnetic particles and streptavidin and mix them evenly in proportion. After mixing, add the blocking solution to obtain magnetic particles coupled with streptavidin, and store them in the first preservation solution to obtain the R1 reagent.

[0039] S3. Take the thrombin antibody and biotin for activation with the second activator and the third activator, respectively, and mix the activated thrombin antibody and the activated biotin in proportion to obtain a biotin-labeled thrombin antibody, Put it in the second preservation solutio...

Example Embodiment

[0050] Example 1

[0051] A kit for determining TAT content includes R1 reagent, R2 reagent and R3 reagent.

[0052] The R1 reagent includes streptavidin-conjugated magnetic particles and a first preservation solution; wherein, the mass ratio of streptavidin-conjugated magnetic particles: streptavidin and activated magnetic particles is 1:5 ; The first preservation solution: 0.9% NaCl, 3% bovine serum albumin, 1% glycerol, 0.5% polyethylene glycol 6000, 0.1% AFP134 antifreeze protein, 0.5% xanthan gum, 0.5% trehalose, 0.02% % Proclin300, the balance is 300 mmol / L Tris-HCl buffer, and the pH is adjusted to 7.3.

[0053] The R2 reagent includes biotin-labeled thrombin antibody and a second preservation solution; wherein, the mass ratio of biotin-labeled thrombin antibody: thrombin antibody and biotin is 1:5; the second preservation solution: 0.9% NaCl, 3 % bovine serum albumin, 0.5% mannitol, 0.5% polyethylene glycol 6000, 0.1% AFP134 antifreeze protein, 0.5% xanthan gum, 0.5%...

Example Embodiment

[0061] Example 2

[0062] A kit for determining TAT content includes R1 reagent, R2 reagent and R3 reagent.

[0063] The R1 reagent includes streptavidin-conjugated magnetic particles and a first preservation solution; wherein, the mass ratio of streptavidin-conjugated magnetic particles: streptavidin and activated magnetic particles is 1:20 ;First preservation solution: 1.0% NaCl, 5.0% bovine serum albumin, 2.0% glycerol, 1.0% polyethylene glycol 6000, 1.0% AFP134 antifreeze protein, 1.0% xanthan gum, 1.0% trehalose, 0.05% % Proclin300, the balance is 600 mmol / L Tris-HCl buffer, and the pH is adjusted to 7.4.

[0064] The R2 reagent includes biotin-labeled thrombin antibody and a second preservation solution; wherein, the mass ratio of biotin-labeled thrombin antibody: thrombin antibody and biotin is 1:20; the second preservation solution: 1% NaCl, 5 % bovine serum albumin, 2% mannitol, 1% polyethylene glycol 6000, 1% AFP134 antifreeze protein, 1% xanthan gum, 1% trehalose,...

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Abstract

The invention discloses a kit for determining TAT content and a preparation method thereof, the kit comprises a reagent R1, a reagent R2 and a reagent R3, the reagent R1 comprises magnetic particles coupled with streptavidin, the reagent R2 comprises a biotin-labeled thrombin antibody, and the reagent R3 comprises an alkaline phosphatase-labeled antithrombin antibody; the method comprises the following steps: uniformly mixing activated magnetic particles and streptavidin to obtain magnetic particles coupled with the streptavidin; uniformly mixing the activated thrombin antibody and biotin to obtain a biotin-labeled thrombin antibody; mixing the activated antithrombin antibody with alkaline phosphatase to obtain an alkaline phosphatase labeled antithrombin antibody; and storing in a preserving fluid to obtain the kit. According to the invention, the amplification effects of streptavidin and biotin are utilized, the magnetic particles are used as a carrier, and the alkaline phosphatase is used as a chemiluminescent marker, so that the sensitivity is high, the specificity is strong, the reagent stability is good, and the result is accurate and reliable.

Description

technical field [0001] The invention relates to the technical field of immunoassays, in particular to a kit for determining TAT content and a preparation method thereof. Background technique [0002] Thrombotic disease refers to a series of diseases caused by the two processes of thrombosis and embolism. It is a common disease and frequently-occurring disease. Its incidence is increasing year by year in the world. Early diagnosis and active prevention can reduce its morbidity and mortality. . The four thrombus items include thrombomodulin, thrombin-antithrombin complex, plasmin-a2 plasmin inhibitor complex, and tissue plasminogen activator-plasminogen activator inhibitor-1 Complex, these 4 substances are obviously increased in the initial stage of coagulation and fibrinolysis system. Among them, the thrombin-antithrombin complex (TAT) is a complex inactive and irreversible covalently linked to form a covalently linked inactive irreversible complex after prothrombin activat...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/573G01N33/543G01N33/58
CPCG01N33/86G01N33/573G01N33/54326G01N33/581G01N2333/974G01N2333/8128G01N2800/226
Inventor 韩学娉
Owner SHENZHEN DYMIND BIOTECH
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