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A kind of avian influenza h9n2 subtype virus strain and application thereof

An avian influenza and virus strain technology, applied in the direction of antiviral agents, virus/phage, antiviral immunoglobulin, etc. Good specific effect

Active Publication Date: 2016-01-13
诗华动保科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Because the bird flu virus mutates very quickly, it brings difficulties to the prevention and control of bird flu, and the research and development cycle of therapeutic drugs is long, which affects the clinical prevention and control effect to a certain extent
Vaccination is the main means of preventing and controlling avian influenza, but its role is greatly limited due to the rapid mutation of avian influenza virus, many epidemic strains, many serological and no cross-immunity

Method used

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  • A kind of avian influenza h9n2 subtype virus strain and application thereof
  • A kind of avian influenza h9n2 subtype virus strain and application thereof
  • A kind of avian influenza h9n2 subtype virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation and Identification of Avian Influenza H9N2 Subtype Virus Strain HF Strain

[0025] 1. Epidemiological investigation

[0026] Cases characterized by severe dyspnea and acute death occurred in a chicken farm in Anhui. 7,800 grass chickens from the age of 20 days died successively within 6 days. More than 300 chickens. After necropsy, it was found that almost all dead chickens had severe hemorrhage of bronchial mucosa, bronchial blockage, full of light yellow cheese-like substance, enlarged kidneys, and dark red congestive necrosis.

[0027] 2. Virus isolation

[0028] Aseptically collect bronchial caseous substances from diseased and dead chickens, as well as their liver, spleen, lungs and other tissues, cut them into pieces and grind them, add normal saline at a ratio of 1:5 to make a suspension, and freeze and thaw three times at -20°C, 3000r / min centrifuged for 15min, the supernatant was filtered with a 0.22μm sterile filter, and the treated ster...

Embodiment 2

[0038] The virulence assay of embodiment 2 avian influenza H9N2 subtype virus strain HF strain

[0039] 1. Determination of intracerebral pathogenicity index (ICPI) of 1-day-old chickens

[0040] Dilute the third-generation SPF chicken embryo allantoic fluid 10 times with sterilized normal saline, filter it through a sterilized filter, and inject 0.05ml into the brain of 10 1-day-old chicks, and inject another 2 chicks in the same way 0.05ml of sterilized saline for diluting the virus. Feed in isolation, observe for 8 days, and record the test results.

[0041] According to the scoring coefficient standard (normal chickens, 0 points; diseased chickens, 1 point; dead chickens, 2 points), the normal, diseased and dead chickens were scored. The test results are shown in Table 2.

[0042] According to the formula for calculating ICPI: ICPI=(number of cases + number of deaths) / total number of observations, the calculation is: ICPI=120 / 80=1.5.

[0043] According to international ...

Embodiment 3

[0062] Immunogenicity Evaluation of Example 3 Avian Influenza H9N2 Subtype Virus Strain HF Strain

[0063] 20 SPF chickens aged 7-10 days were used, among which 10 chickens were subcutaneously inoculated with HF strain allantoic fluid diluted 1:10, and the other 10 were subcutaneously inoculated with normal saline as a control, and were isolated and raised separately. After 21 days, blood was collected to separate serum. Use standard H9 antigen for HI test. The results are shown in Table 6. The results showed that the HF strain had good immunogenicity.

[0064] Immunogenicity of table 6HF strains

[0065]

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Abstract

The invention provides an avian influenza H9N2 subtype virus strain with the preservation number of CGMCC No.6757. The avian influenza H9N2 subtype virus strain provided by the invention has favorable specificity and immunogenicity, the erythrocyte agglutination effect of allantoic fluid can not be inhibited by anti-NDV (Newcastle Disease Virus) positive serum, anti-EDS76 positive serum, anti-M41 positive serum, anti-H5 subtype avian influenza virus positive serum and anti-H7 subtype avian influenza virus positive serum, but can be inhibited by H9 subtype avian influenza virus positive serum, and immunized chicken can generate an HI (Hemagglutination Inbition) antibody which is specific for H9. The avian influenza H9N2 subtype virus strain provided by the invention can be used as a vaccine strain for preventing H9 subtype avian influenza of birds, and can be used for authenticating avian influenza viruses and researching epidemiology so as to have favorable market application prospects.

Description

technical field [0001] The invention belongs to the field of microbial viruses, and in particular relates to an avian influenza H9N2 subtype virus strain and application thereof. Background technique [0002] Avian influenza (Avian influenza, AI) is a highly infectious disease caused by type A avian influenza virus (Avian influenza virus, AIV), which seriously endangers the poultry industry. Avian influenza viruses belong to the family Orthomyxoviridae, the genus Influenzavirus. Hemagglutinin (HA) and neuraminidase (NA) are the two main protective antigens of the virus. According to the antigenic differences of HA and NA, influenza A viruses can be divided into different subtypes. So far, 16 types of HA and 10 types of NA have been found for type A avian influenza virus, named after H1-H16 and N1-N10 respectively, and there is no cross-reaction between different H antigens or N antigens. The amino acid sequence homology of each subtype of hemagglutinin is less than 70%, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C07K16/10C07K16/06A61K39/145A61P31/16G01N33/569C12R1/93
Inventor 张坦郑杰张红孙丰廷杨国良王亚丽张凌云张发明张洪
Owner 诗华动保科技(北京)有限公司
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