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Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof

A hepatitis A virus and chemiluminescence immunological technology, applied in the field of immunoassay medicine, can solve problems such as unseen use, and achieve the effects of ensuring sensitivity, facilitating large-scale production, and reducing gray area values.

Inactive Publication Date: 2009-09-16
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, chemiluminescent immunoassay technology is widely used in in vitro diagnostic immunoassay reagents, but the application of directly coated antigen combined with chemiluminescent immunoassay technology has not been used in the application of hepatitis A virus IgM antibody immunoassay products

Method used

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  • Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof
  • Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof
  • Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of the hepatitis A virus IgM antibody chemiluminescent immunoassay assay kit of the present invention

[0029] In the research process of the present invention, the inventor of the present invention firstly carried out screening experiment and quality appraisal to the raw material used, comprise the activity of antibody marker and coated antibody, carrier (such as opaque white microwell plate) Adsorption performance and variability, HRP activity, luminescence intensity and luminescence duration of chemiluminescent substrates, etc. Then the coating method was studied, experiments were carried out with different coating buffers and protection solutions, the most suitable coating buffer and protection solutions were selected, and the best concentration conditions were found through experiments with different coating concentrations of antibodies. There are different methods for HRP labeling. Through repeated exploration and comparative experiments, a s...

Embodiment 2～3

[0085] Examples 2-3 Preparation of Hepatitis A virus IgM antibody chemiluminescent immunoassay assay kit of the present invention

[0086] Except that the hepatitis A virus antibody was labeled with alkaline phosphatase, AMPPD was used as the chemiluminescent substrate, plastic beads and plastic tubes were used as the carrier, the pH value of the coating solution was 4.5, and the pH value of the blocking solution was 7.5. The hepatitis A virus IgM antibody chemiluminescent immunoassay assay kit was prepared in the same manner as in Example 1.

Embodiment 4

[0087] Example 4 Preparation of the hepatitis A virus IgM antibody chemiluminescence immunoassay assay kit of the present invention

[0088] Except that magnetic particles were used as the solid phase carrier, and the magnetic particle anti-human-μ chain antibody solid phase carrier was prepared by the glutaraldehyde coupling method, the rest were prepared by the same method as in Example 1. The hepatitis A virus IgM antibody chemiluminescence immunoassay Analytical assay kits.

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Abstract

The invention relates to the field of immunoassay medical science, and in particular provides a chemoluminescent immunoassay kit of a hepatitis A virus IgM antibody and a preparation method thereof. The kit comprises: 1) negative and positive control varieties of the hepatitis A virus IgM antibody; 2) a dermatate solid phase carrier; 3) a hepatitis A virus antigen liquid; 4) an enzyme label; 5) a chemoluminescent substrate; and 6) a condensed washing solution. Furthermore, the method for preparing the kit comprises the following steps: 1) preparing control varieties from negative and positive serum of the hepatitis A virus IgM antibody; 2) dermatating the solid phase carrier by an anti-human-mu chain antibody (monoclonal antibody or polyclonal antibody); 3) preparing an antigen liquid from the hepatitis A virus; 4) labeling the hepatitis A virus antibody (the monoclonal antibody or the polyclonal antibody) by enzyme; 5) preparing the chemoluminescent substrate solution; 6) preparing the condensed washing solution; 7) sub-packaging the negative and positive control varieties of the hepatitis A virus IgM antibody, the hepatitis A virus antigen liquid, the enzyme label, the chemoluminescent substrate solution and the condensed washing solution; and 8) assembling the substances into the finished products. The kit has the advantages of simpleness, convenience, quickness, sensitiveness, stability, and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a hepatitis A virus IgM antibody chemiluminescent immunoassay kit and a preparation method thereof. Background technique [0002] Hepatitis A is an infectious disease mainly caused by liver damage caused by hepatitis A virus (HAV) transmitted through the fecal-oral route. It is an RNA virus belonging to Picornaviridae, enterovirus type 72, spherical in diameter with a diameter of about 27nm, and composed of 32 shell particles with a symmetrical 20-hedron nucleocapsid containing linear single-stranded RNA. HAV has 4 main polypeptides, which are the main antigenic polypeptides constituting the viral capsid protein, which induce neutralizing antibodies. HAV exists in the patient's blood, feces and liver cytoplasm. Anti-HAV1gM antibodies in serum appeared soon after infection and reached a peak in about 2 weeks. Then it gradually decreased and disappeare...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/569G01N21/76G01N33/577G01N33/543
CPCY02A50/30
Inventor 杨俊峰应希堂胡国茂郑金来于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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