H5 subtype waterfowl flu virus hemagglutinin antibody indirect ELISA kit

A detection kit and antibody detection technology, applied in the biological field, can solve problems such as the confusion of influenza antibody detection methods, and achieve the effects of easy large-scale promotion and application, good safety, and strong specificity

Inactive Publication Date: 2009-01-14
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the particularity of the species of waterfowl, the detection method of its influenza antibody has always been a problem that perplexes the evaluation of the immune effect of waterfowl vaccines and the diagnosis of waterfowl influenza.

Method used

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  • H5 subtype waterfowl flu virus hemagglutinin antibody indirect ELISA kit
  • H5 subtype waterfowl flu virus hemagglutinin antibody indirect ELISA kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] According to technical scheme of the present invention, concrete preparation is as follows:

[0028] (1) Preparation of AIV / H5 recombinant HA protein

[0029]Using RT-PCR technology to amplify part of the hemagglutinin protein gene sequence (1020bp fragment at the 5' end of the HA gene) from the H5 subtype waterfowl influenza genome (see Figure 3), insert it into the downstream of the prokaryotic expression vector PET28a to construct PET -H5 / HA expression vector, and transform Escherichia coli BL21(DE3) with calcium chloride method, induce expression with 1mM IPTG, lyse the bacteria with ultrasonic waves, and purify the expressed protein (HA) using a commercial Ni-NTA affinity chromatography column, Obtain part of the hemagglutinin protein of waterfowl influenza virus ( figure 2 ), the protein retains the antigenicity of most of the hemagglutinin protein, and can specifically bind to the corresponding subtype hemagglutinin antibody of waterfowl influenza virus ( fig...

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Abstract

This invention discloses indirect ELISA kit for hemagglutinin antibody of H5 subtype water bird flu virus, which contains antibody test board, enzyme conjugate work solution, positive control, negative control, sample diluent, concentrated detergent, color development solution A, color development solution B, and stop solution, wherein the test board is dismountable 96 holes microtiter plate of peridium water bird flu H5 subtype hemagglutinin protein (HA), the enzyme conjugate work solution is HRP rabbit antibody against water bird IgG polyclonal antibody, positive control is H5 subtype water bird flu standard positive serum, negative control is water bird standard negative serum.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rapid diagnosis method and equipment for a virus antibody of waterfowl, in particular to an indirect ELISA detection kit for H5 subtype waterfowl influenza virus (AIV) hemagglutinin antibody. Background technique [0002] Avian influenza (avian influenza), the full name of avian influenza virus infection, is an infectious disease of poultry caused by type A influenza virus of Orthomyxoviridae and Influenza virus, from respiratory system infection to sepsis, etc., poultry The fatality rate after infection is high. Bird flu was first discovered in Italy by Perroncito in 1878. Its pathogen was discovered for the first time in 1900, and it was only confirmed by serology in 1955 that it belonged to type A (A) influenza virus. Since then, avian influenza viruses have been prevalent in poultry around the world, with varying degrees of impact. In particular, the outbreak of bird flu in Ho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/531G01N21/31
Inventor 周继勇陈洪勋申会刚
Owner ZHEJIANG UNIV
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