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

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Problems solved by technology

However, this method is cumbersome to detect, has poo

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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

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Example Embodiment

[0051] Example 1

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

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

[0054] (2) Preparation of R2: The preparation of 150nm latex microsphere system includes the following steps: using 100mL of EDC solution with a mass volume percentage of 2% and 100mL of 150nm latex microspheres with a mass volume percentage of 5% at 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 pH 6.5-7.0 MES buffer; then add 0.005% by mass and volume of mouse anti-human von Willebrand factor monomer The cloned antibody was mixed and incubated at 37°C for 5 hours. After incubation, it was washed 5 tim...

Example Embodiment

[0057] Example 2

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

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

[0060] (2) Preparation of R2: The preparation of 150nm latex microspheres system includes the following steps: using 100mL of EDC solution with a mass volume percentage of 1% and 100mL of 150nm latex microspheres with a mass volume percentage of 15% at 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; then add mouse anti-human von Willebrand factor monomer with a mass volume percentage of 0.001% The cloned antibody was mixed and incubated at 37°C for 5 hours, and then wash...

Example Embodiment

[0063] Example 3

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

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

[0066] (2) Preparation of R2: The preparation of 150nm latex microsphere system includes the following steps: use 100mL of EDC solution with a mass volume percentage of 10% and 100mL of 150nm latex microspheres with a mass volume percentage of 1% at 37°C 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 pH 6.5-7.0 MES buffer; then add the mouse anti-human von Willebrand factor monomer with a mass volume percentage of 0.05% The cloned antibody was mixed and incubated at 37°C for 5 hours. After incubation, washed 5 times with pH 6.5-7.0 MES buff...

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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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