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Vascular hemophilia factor detection reagent, preparation method and application thereof

A hemophilia factor and detection reagent technology, applied in the field of clinical medical detection, can solve the problems of lack of reliability, cumbersome detection, low repeatability, etc., and achieve the effects of improving coupling efficiency, accurate test results, and stable detection

Inactive Publication Date: 2018-02-09
北京众驰伟业科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is cumbersome to detect, has poor stability, low repeatability, and lacks reliability.

Method used

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  • Vascular hemophilia factor detection reagent, preparation method and application thereof
  • Vascular hemophilia factor detection reagent, preparation method and application thereof
  • Vascular hemophilia factor detection reagent, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0052] The preparation method of the detection reagent of von Willebrand factor in this embodiment comprises the following steps:

[0053] (1) Preparation of R1: first prepare the MES buffer solution with a concentration of 50 mM, then add 0.5% NaCl by mass volume percentage and 0.08% Triton X-100 by mass volume percentage, adjust the pH to 7.2 after mixing .

[0054] (2) Preparation of R2: The preparation of the 150nm latex microsphere system comprises the following steps: 100mL of 150nm latex microspheres 100mL of 150nm latex microspheres with a mass volume percentage of 2% and a mass volume percentage of 5% with a 37°C constant temperature shaker Shake for 25 minutes, centrifuge the activated microspheres at 3000 rpm for 10 minutes, discard the supernatant, and wash 5 times with pH6.5-7.0 MES buffer; The cloned antibody mixture was incubated at 37°C for 5 hours, and washed 5 times with MES buffer solution of pH 6.5-7.0 after incubation; then added MES buffer solution of pH...

Embodiment 2

[0058] The preparation method of the detection reagent of von Willebrand factor in this embodiment comprises the following steps:

[0059] (1) Preparation of R1: first prepare the MES buffer solution with a concentration of 10 mM, then add 0.1% NaCl by mass volume percentage and 0.01% Triton X-100 by mass volume percentage, adjust the pH to 6.8 after mixing .

[0060] (2) Preparation of R2: The preparation of the 150nm latex microsphere system comprises the following steps: 100mL of 150nm latex microspheres with a mass volume percentage of 1% and 150nm latex microspheres of 15% by mass volume are placed on a 37°C constant temperature shaker Shake for 30 minutes, centrifuge the activated microspheres at 3000 rpm for 10 minutes, discard the supernatant, and wash 4 times with pH6.5-7.0 MES buffer; The cloned antibody mixture was incubated at 37°C for 5 hours, and washed once with MES buffer of pH 6.5-7.0 after incubation; then MES buffer of pH 6.5-7.0 containing 0.1% bovine seru...

Embodiment 3

[0064] The preparation method of the detection reagent of von Willebrand factor in this embodiment comprises the following steps:

[0065] (1) Preparation of R1: first prepare the MES buffer solution with a concentration of 200 mM, then add 3% NaCl by mass volume percentage and 1% Triton X-100 by mass volume percentage, adjust the pH to 7.4 after mixing .

[0066] (2) Preparation of R2: The preparation of the 150nm latex microsphere system comprises the following steps: 100mL of 100mL of 150nm latex microspheres with a mass volume percentage of 10% by mass volume percentage and 100mL of 150nm latex microspheres by mass volume percentage at 37°C on a constant temperature shaker Shake for 20 minutes, centrifuge the activated microspheres at 3000 rpm for 10 minutes, discard the supernatant, and wash 5 times with pH6.5-7.0 MES buffer; The cloned antibody mixture was incubated at 37°C for 5 hours, and washed 5 times with MES buffer solution of pH 6.5-7.0 after incubation; then, ME...

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Abstract

The invention provides a vascular hemophilia factor detection reagent, a preparation method and application thereof. The detection reagent includes two latex microsphere systems with different particle sizes. The vascular hemophilia factor detection reagent prepared through the preparation method is high in repeatability and accurate and rapid in detection, limiting influencing factors are fewer,results are reliable, and the detection reagent is low in cost and beneficial to clinical medical applications.

Description

technical field [0001] The invention belongs to the field of clinical medical detection, and in particular relates to a detection reagent for von Willebrand factor and its preparation method and application. Background technique [0002] Coagulation tests are of great significance to the diagnosis of diseases in various clinical departments. In addition to the screening and diagnosis of bleeding diseases, they are also used for the identification of von Willebrand factor defects and the determination of hemophilia. [0003] Von Willebrand factor (von Willebrand factor, vWF) is synthesized and secreted by vascular endothelial cells and megakaryocytes, and is a glycoprotein present in plasma, endothelial cell surface and platelet α granules. In the body, vWF mainly exists in the form of dimers connected by disulfide bonds, but it can also be polymerized into multimers with a molecular weight of one million to more than ten million. Its defect will cause von Willebrand disease ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/544G01N33/577
CPCG01N33/6863G01N33/54313G01N33/544G01N33/577G01N2333/4728G01N2333/755G01N2800/224
Inventor 杨军京
Owner 北京众驰伟业科技发展有限公司
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