Enzyme-linked immunologic adsorption test method for identification and diagnosis of chicken infected with avian influenza

Abstract

The invention relates to the enzyme combined immune absorbing testing for diagnosing bird flu affected chicks. It expands H9N2 bird flu virus, acquiring target section to connect with the carrier to restructure the expressing particle, injecting into bacillus coli expression to get a conjugated protein which is purified to get H9N2 bird flu virus NS1 protein to be antibody wrapping enzyme reaction plate, to build the antibody level to be reacted to inspect of the serum, getting the marginal value and least sample number of the enzyme combined immune adhesive test through statistics, and finally based on the average OD value to identify the bird flu affection status of the overall poultry. Through overall diagnosis, it alleviates nonspecific reaction, provides scientific proof for timely diagnosis and prevention of bird flu.

Description

technical field [0001] The invention relates to a method in the technical field of bioengineering, in particular to an enzyme-linked immunosorbent assay method for differential diagnosis of bird flu-infected chicken flocks. Background technique [0002] Bird flu, especially HPAI (highly pathogenic bird flu), has great harm to the poultry farming industry. At present, one of the methods to effectively control avian influenza is vaccination. An index to effectively evaluate the immune effect is to detect the level of antibodies after immunization. Therefore, it is very important to effectively distinguish the antibodies produced by natural infection from those produced by immunization. Traditional serological methods for avian influenza diagnosis include HA-HI (hemagglutination and hemagglutination inhibition test), AGP (agar immunodiffusion), microneutralization and NIF (neuraminidase inhibition test). Antibodies can be produced no matter whether they are vaccinated or infec...

AI Technical Summary

Problems solved by technology

However, on the one hand, NS1 protein is a weak antigen, which is not easy to stimulate the body to produce high-titer antibodies

That is, naturally infected chickens may not produce high antibody titers at different individual levels; For the production of NS1 protein, there is NS1 protein in the oil emulsion inactivated vaccine prepared by using chicken embryo allantoic fluid, so there is likely to be a certain amount of antibody against NS1 protein in immunized chicken flocks, especially in chicken flocks that have been immunized many times. That is to say, at the individual level, the level of NS1 antibody in chickens naturally infected with avian influenza virus may not be high, or the level of NS1 antibody in immunized chickens may be relatively high, so the differential diagnosis of avian influenza infection at the individual level is often difficult. big deviation

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