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

An enzyme-linked immunosorbent adsorption and differential diagnosis technology, applied in the field of bioengineering, can solve problems such as deviation, weak antigen, and low

Inactive Publication Date: 2007-07-11
SHANGHAI JIAO TONG UNIV
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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 agai...

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

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Embodiment Construction

[0021] The embodiments of the present invention are described in detail below: the present embodiment is implemented under the premise of the technical solution of the present invention, and detailed implementation and specific operation process are provided, but the protection scope of the present invention is not limited to the following implementation example.

[0022] The experimental method that does not indicate specific condition in the embodiment, usually according to conventional conditions, such as Sambrook etc. in " Molecular Cloning Laboratory Handbook " (New York Cold Spring Harbor Laboratory Press, 1989) described conditions, or according to manufacturer's suggested conditions.

[0023] In this example, the H9N2 avian influenza virus was first amplified by reverse transcription polymerase chain reaction to obtain the target fragment, and then the target fragment was connected to the vector pET-32a(+) plasmid to obtain a recombinant expression plasmid, which was t...

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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...

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Application Information

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IPC IPC(8): G01N33/569G01N33/543
Inventor 孙建和王琰陆承平严亚贤
Owner SHANGHAI JIAO TONG UNIV
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