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Reagent for detecting Von Willebrand factor and preparation method and application thereof

A technology for detecting reagents and factors, applied to a kit for detecting von Willebrand factor in human serum and the field of preparation thereof, can solve the problems of inconvenient detection process, small linear range and the like, and achieve rapid and thorough reaction and good stability , It is not easy to be interfered by human operation and external factors

Inactive Publication Date: 2014-04-30
SHANGHAI VASCUTECH DIAGNOSIS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reason is that the linear range of detection is small, which brings a lot of inconvenience to the detection process

Method used

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  • Reagent for detecting Von Willebrand factor and preparation method and application thereof
  • Reagent for detecting Von Willebrand factor and preparation method and application thereof
  • Reagent for detecting Von Willebrand factor and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] a) Preparation of Reagent 1

[0069] Reagent 1 is an antibody latex reagent for anti-von Willebrand factor obtained by reacting latex particles containing carboxyl groups on the surface with amino groups of antibodies under the action of a binding agent; the specific preparation method is as follows:

[0070] Prepare 0.05M KH 2 PO 4 For pH 4.5 activation buffer, carboxy-modified latex particles with a particle size of 500nm were washed three times with pure water and then suspended in 10ml of activation buffer. The final concentration of latex particles was 1% by mass and volume in g / ml. The acidic pH of the activation buffer is to ensure that the carboxyl group is in a stable state of carboxyl coordination, which is conducive to the reaction.

[0071] Then freshly prepare 2% carbodiimide, take 10ml and add it into latex particles of equal volume, stir and react at room temperature (18-25° C.) for 15 minutes; thereby activate the latex particles.

[0072] Prepare 0.2...

Embodiment 2

[0079] The difference between this example and Example 1 is only that the particle size of the latex particles is 1000 nm, and other operations and proportions are consistent with Example 1.

[0080] The specificity, sensitivity and linear range of the obtained kit are shown in the following experiments.

[0081] Detection of specificity, sensitivity and linear range of the kit

[0082] 1. The detection operation procedure on the automatic detection instrument is as follows:

[0083] (1) Freshly drawn venous blood was mixed with citrate anticoagulant solution at a ratio of 9:1, centrifuged, and separated to obtain plasma;

[0084] (2) Set parameters on the automatic detector: reaction temperature 37°C; analysis method: 2-point endpoint method; main wavelength: 570nm, secondary wavelength: 800nm; sample volume / reagent 1 / reagent 2: 20μl / 100μl / 100μl; Reaction direction: up;

[0085] (3) Select von Willebrand factor standard, use the application solution for serial dilution: S7...

Embodiment 3

[0103] a) Preparation of Reagent 1

[0104] Reagent 1 is an antibody latex reagent for anti-von Willebrand factor obtained by reacting latex particles containing carboxyl groups on the surface with amino groups of antibodies under the action of a binding agent; the specific preparation method is as follows:

[0105] Prepare 0.0SM KH 2 PO 4 For pH 4.5 activation buffer, carboxy-modified latex particles with a particle size of 500nm were washed three times with pure water and then suspended in 10ml of activation buffer. The final concentration of latex particles was 1% by mass and volume in g / ml. The acidic pH of the activation buffer is to ensure that the carboxyl group is in a stable state of carboxyl coordination, which is conducive to the reaction.

[0106] Then freshly prepare 2% carbodiimide, take 10ml and add it into latex particles of equal volume, stir and react at room temperature (18-25° C.) for 10 minutes; thereby activate the latex particles.

[0107] Prepare 0.2...

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Abstract

A reagent for detecting von Willebrand factor. The detection reagent applies latex enhanced immunoturbidimetry method, and comprises a reagent 1, a reagent 2 and a von Willebrand factor standard substance or a standard curve. The reagent 1 is an anti-von Willebrand factor antibody latex reagent, which is formed by combining the anti-von Willebrand factor and latex particles; the reagent 2 is an application liquid; the von Willebrand factor standard substance is a reaction buffer containing von Willebrand factors with different concentrations; and the standard curve is a standard curve of the content of von Willebrand factor in human body. Compared with the prior art, the reagent for detecting von Willebrand factor provided by the invention realizes rapid detection of von Willebrand factor, and has high sensitivity and reliable repeatability, and can be applied to automatic analytical instruments.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a kit for detecting von Willebrand Factor (vWF, von Willebrand Factor) in human serum and a preparation method thereof. Background technique [0002] Von Willebrand Factor (vWF, von Willebrand Factor), also known as von Willebrand Factor, is encoded by the vWF gene on chromosome 12 and is a large molecular weight multi-subunit glycoprotein composed of multiple A 220kD subunit. The degree of polymerization of the final product varies, and the highest molecular weight can be as high as 20MD. After assembly, it is stored in α-granules of platelets and Weibel-Palade bodies in vascular endothelium; it is a multifunctional and multivalent large adhesion glycoprotein present in blood. vWF factor has an extremely important physiological role in the process of hemostasis and thrombus formation: on the one hand, it regulates the platelet adhesion response, and re...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N33/54393G01N2800/224G01N2800/226G01N2800/32Y10S436/811
Inventor 赵铁铭李小林王枕亚喜庆
Owner SHANGHAI VASCUTECH DIAGNOSIS CO LTD
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