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H9N2 avian influenza virus strain, and prepared inactivated vaccine and application thereof

A bird flu virus and bird flu technology, applied in antiviral agents, viruses/bacteriophages, biochemical equipment and methods, etc., can solve problems such as high cost, chicken-derived potential disease infection, large batch-to-batch difference in product quality, etc., to achieve High adaptability, rapid increase in antibody levels, and the effect of preventing infection

Inactive Publication Date: 2015-01-28
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the commercially available avian influenza vaccines are chicken embryo tissue seedlings. Compared with cell seedlings, they have disadvantages such as high cost, large batch-to-batch variation in product quality, and prone to chicken-derived potential disease infection.

Method used

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  • H9N2 avian influenza virus strain, and prepared inactivated vaccine and application thereof
  • H9N2 avian influenza virus strain, and prepared inactivated vaccine and application thereof
  • H9N2 avian influenza virus strain, and prepared inactivated vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Isolation and Identification of H9N2 Subtype Avian Influenza Virus NJ13-8 Strain

[0037] 1. Experimental method

[0038] 1.1 Isolation of virus

[0039] In 2013, the lungs and livers were collected from a chicken farm in Nanjing, Jiangsu Province, and sent for inspection. The lungs and livers were cut into pieces, added with sterilized saline at a ratio of 1:5, ground in a sterile mortar, frozen and thawed three times repeatedly, and subjected to 3000r / min centrifuged for 15min, and the supernatant was taken for later use.

[0040] The supernatant of the above-mentioned disease material was sterilized through a 0.22 μm disposable filter, and inoculated with 10-day-old SPF chicken embryos through the allantoic cavity, with 0.1 ml for each chicken embryo, and a total of 5 chicken embryos were inoculated. Incubate at 37°C; discard dead embryos within 24 hours, aseptically collect the allantoic fluid of chicken embryos that died within 24-120 hours, and store ...

experiment example 1

[0067] Experimental example 1 Pathogenicity assay of isolated virus NJ13-8 strain and its proliferation in MDCK cells 1, experimental method

[0068] 1.1 Pathogenicity determination of isolated virus NJ13-8 strain

[0069] Embryo median infectious dose (EID) 50 ), intracerebral pathogenicity index (ICPI) and intravenous inoculation index (IVPI) test to detect the pathogenicity of the virus NJ13-8 strain that embodiment 1 isolates.

[0070] 1.1.1 Virus EIDs 50 determination

[0071] Make the isolated virus with sterilized saline for 10 -6 ~10 -9 Dilution, and set up a blank control group, inoculate 5 10-day-old SPF chicken embryos at each dilution, discard 24h dead embryos, freeze the chicken embryos overnight at 4°C 120h after inoculation, harvest chicken embryo allantoic fluid, and measure its HA efficacy Valence, and refer to the Reed-Muench method to calculate the virus EID 50 .

[0072] 1.1.2 Determination of intracerebral pathogenicity index (ICPI)

[0073] Take 1...

experiment example 2

[0110] Preparation of experimental example 2 inactivated vaccine and immune protection test

[0111] 1. Experimental method

[0112] 1.1 Amplification of virus

[0113] The virus NJ13-8 strain that embodiment 1 isolates is respectively through MDCK cell culture amplification (collecting cell culture supernatant) and inoculation SPF chicken embryo amplification (collecting allantoic fluid), after the virus liquid collection of above amplification is evenly packed , stored at -70°C for later use.

[0114] 1.2 Preparation of inactivated vaccine

[0115] Take the two virus liquids amplified by MDCK cells and chicken embryos, centrifuge at 5000r / min for 30min, take the supernatant, add formalin solution to the final concentration of 0.2% of the total amount of the virus liquid supernatant, and store at 37°C After 24 hours of inactivation, after the sterility test, the inactivated virus was mixed with imported white oil adjuvant according to the volume ratio (1:3), emulsified and...

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Abstract

The invention discloses an H9N2 avian influenza virus strain, a prepared inactivated vaccine and application thereof. The microbial preservation number of the H9N2 avian influenza virus strain NJ13-8 is CGMCC No.9325. The invention also discloses a method for preparing the H9N2 avian influenza inactivated vaccine. The method comprises the steps of amplifying virus strain NJ13-8, collecting virus solution, centrifuging to obtain supernatant, adding inactivators into the supernatant of the virus solution for inactivation, adding adjuvants into the inactivated supernatant of the virus solution, mixing uniformly, and emulsifying. The separated virus strain NJ13-8 has high adaptation to MDCK cells, cell culture shows that the virus content is higher than that of the strain obtained through chick-embryo culture, the production period is short, the cost is low, and mass production is facilitated; and compared with the vaccine prepared by embryo virus, the inactivated vaccine has high valence of antibody, the antibody level can be raised rapidly, and the infection of the H9N2 avian influenza virus can be effectively prevented.

Description

technical field [0001] The present invention relates to avian influenza virus strains, in particular to an isolated H9N2 subtype avian influenza virus strain, and also relates to the application of the strain in the preparation of a vaccine against avian influenza, which belongs to the isolation and application of H9N2 subtype avian influenza virus technology field. Background technique [0002] Avian influenza, referred to as avian influenza (Avain Influenza, AI), is an infection and / or disease syndrome of poultry (including poultry and wild fowl) caused by type A influenza virus, mainly causing systemic or respiratory system infection in poultry. The disease was established as a Class A infectious disease by the World Organization for Animal Health. According to the different pathogenicity of avian influenza, avian influenza can be divided into highly pathogenic avian influenza, low pathogenic avian influenza and apathogenic avian influenza. [0003] H9N2 subtype avian i...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/145A61P31/16C12R1/93
Inventor 刘宇卓李银黄欣梅赵冬敏韩凯凯杨婧滕颖
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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