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H9 subtype avian influenza virus Vero cell adapted strain and application thereof

A technology of avian influenza virus and adaptive strains, applied in antiviral agents, viruses/bacteriophages, medical preparations containing active ingredients, etc., can solve problems such as infection, large batch-to-batch difference in product quality, and high cost, so as to reduce production costs, The effect of broad application prospects

Active Publication Date: 2012-07-18
哈药集团生物疫苗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Vaccination is the most effective way to prevent the disease. At present, most of the avian influenza vaccines available on the market are chicken embryo tissue vaccines. And it is easy to produce chicken-derived potential disease infection and other shortcomings

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Obtaining of H9 subtype avian influenza virus VERO cell-adapted strain AIV HY strain

[0032] 1 Isolation of the original virus strain

[0033] The disease material came from a farm in the suburbs of Shuangcheng City, Heilongjiang Province. Tracheal swabs and cloacal swabs were collected from dying chickens, treated with 2000 units of penicillin and streptomycin solution, inoculated with chicken embryos, and collected 48h-78h chicken embryo allantoic fluid . The hemagglutination test was carried out to the harvested chicken embryo fluid, and the hemagglutination value of HA was 9log2.

[0034] 2 Hemagglutination inhibition (HI) test of virus isolates

[0035] HI test was carried out with H5, H7 and H9 subtype AI standard positive serum, ND, EDS-76 virus standard positive serum and HA positive chicken embryo allantoic fluid respectively. The results showed that the standard positive seroreaction with H5, H7, ND and EDS-76 virus was negative, and the standard...

Embodiment 2

[0040] Embodiment 2 utilizes H9 subtype avian influenza virus VERO cell adaptation strain AIV HY strain to produce influenza virus inactivated vaccine

[0041] 1. Preparation of immobilized trypsin by chitosan nanoparticle embedding method

[0042] (1) Weigh chitosan with a degree of deacetylation of 85%, dissolve it with 1% acetic acid to make a chitosan solution with a concentration of 0.5-2 mg / mL, and pass it through a 0.22 μm filter. Weigh trypsin, dissolve it in deionized water to make a trypsin solution with a concentration of 500 μg / mL, and pass it through a 0.22 μm filter. Weigh sodium tripolyphosphate, dissolve it with deionized water to make a sodium tripolyphosphate solution with a concentration of 1.0-2.0 mg / mL, and pass it through a 0.22 μm filter.

[0043] (2) Take 5 mL of chitosan solution, add 2.5-7.5 mL of 500 μg / mL trypsin solution dropwise, and stir magnetically for 3 minutes to obtain solution A;

[0044] (3) Solution A was magnetically stirred at 900-130...

Embodiment 3

[0063] Embodiment 3 utilizes the comparison of the vaccine prepared by cytotoxicity and tissue toxicity

[0064] Produced according to the method of Example 2, the vaccine produced by utilizing the domesticated H9 subtype avian influenza virus VERO cell-adapted strain AIV HY strain (cytotoxicity) of the present invention and produced according to the method of Experimental Example 1, utilizing undomesticated H9 The vaccines produced by subtype avian influenza virus strains (histotoxicity) were compared. In order to verify the characteristics of cytotoxicity, 10 batches of histotoxicity and cytotoxicity were prepared respectively, and the relevant technical indicators were determined. Now some of the indicators are compared. The results are summarized below, as described in Tables 1-5:

[0065] Table 1 Comparison of agglutination values ​​of two kinds of toxic red blood cells

[0066] batch 1 2 3 4 5 6 7 8 9 10 tissue poison 1∶128...

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Abstract

The invention discloses a H9 subtype avian influenza virus Vero cell adapted strain and application of the Vero cell adapted strain in preparing of H9 subtype avian influenza vaccines. When being applied to Vero cells, viral multiplication speed and yield of the adapted strain are all superior to the existing avian influenza virus cultured through chicken embryos and superior to viral strains of non-naturalized H9 subtype avian influenza virus. The H9 subtype avian influenza virus Vero cell adapted strain can replace the chicken embryos to massively produce influenza viruses fast and has wideapplication prospect.

Description

technical field [0001] The invention relates to a cell strain capable of stably proliferating in Vero cells and its application, in particular to an H9 subtype avian influenza virus Vero cell-adapted strain and its application. It belongs to the field of vaccine preparation. Background technique [0002] Avian influenza (abbreviated as avian influenza, AI) is an infection and / or disease syndrome of poultry caused by type A influenza virus. The pathogen that causes AI is called avian influenza virus. [0003] According to the disease classification standard of the World Organization for Animal Health (OIE), highly pathogenic avian influenza is a type A disease, and my country also lists it as a type of infectious disease, with fast transmission speed, high morbidity and mortality; AIV with low virulence can cause subclinical symptoms, such as decreased egg production, respiratory system or systemic infection, and various symptoms of varying degrees. Chicken flocks are infec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/145A61P31/16C12R1/93
Inventor 丁国杰张杨孙德君吴金姜力张明艳
Owner 哈药集团生物疫苗有限公司
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