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Duck-sourced H4N6 subtype avian influenza virus strain and application thereof

A technology of influenza A virus and influenza virus, which is applied in the field of animal virology, can solve problems such as the decline in egg production and the loss of poultry farming

Inactive Publication Date: 2014-02-12
SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these viruses are mostly low pathogenic to animals, they can cause losses to the poultry industry by reducing egg production

Method used

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  • Duck-sourced H4N6 subtype avian influenza virus strain and application thereof
  • Duck-sourced H4N6 subtype avian influenza virus strain and application thereof
  • Duck-sourced H4N6 subtype avian influenza virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1H4

[0035] Isolation and preservation of embodiment 1H4N6 subtype avian influenza virus

[0036] 1. Isolation of H4N6 subtype avian influenza virus

[0037] 20 swabs of throat and cloaca of healthy ducks in a duck farm in Shanghai, my country were collected for isolation, culture and identification. One virus strain capable of agglutinating chicken erythrocytes was isolated through chicken embryo culture, and identified as H4N6 subtype avian influenza virus, as follows.

[0038] (1) Virus isolation and culture: collect swabs of duck throat and cloaca, store them in PBS buffer, and transport them to the laboratory within 4 hours of cold chain. Duck throat and cloaca cotton swab samples were processed and cultured according to the method of GB / T18936-2003. The samples were treated with penicillin (3000IU / mL) and streptomycin (3mg / mL) at 4°C for 1 hour, and the cotton swabs were fully twisted and wrung out. Afterwards, the swab was discarded, the sample solution was centrifuged at 10...

Embodiment 2

[0070] The preparation of embodiment 2 virus strains

[0071] The preservation number is the avian influenza virus strain of CGMCC NO.7606 with 0.2mL / piece (10 4 ~10 5 EID 50 / mL) to inoculate 9-10-day-old SPF chicken embryos, place the inoculated chicken embryos in an incubator for 48-96 hours, discard the dead chicken embryos within 24 hours, and collect the allantoic fluid of infected chicken embryos. . The cultivation conditions are:

Embodiment 3

[0072] The preparation of embodiment 3 vaccines

[0073] The virus strain of the invention is used to prepare H4N6 subtype avian influenza virus vaccine.

[0074] 1. Vaccine preparation

[0075] (1) Centrifuge the prepared DK / SH / Y20 / 2006 strain virus liquid at 5000×g for 10 min, and take the supernatant.

[0076] (2) Add formaldehyde to the supernatant so that the volume percentage of formaldehyde is 0.1%, inactivate at 37°C for 24 hours (start timing when the temperature in the bottle reaches 37°C), shake 3-4 times during the period, and obtain inactivated virus fluid. Take the inactivated virus liquid, inoculate 9-10 day-old SPF chicken embryos, pass the inspection and set aside.

[0077] (3) emulsification

[0078] Oil phase preparation: 94 parts by volume of white oil for injection (purchased from Hangzhou Oil Refinery) and 6 volume parts of Spin-80 (purchased from Shanghai Dazhong Pharmaceutical Co., Ltd.), mixed well, and then added aluminum stearate (purchased from Sh...

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Abstract

The invention discloses a duck-sourced H4N6 subtype avian influenza virus strain and an application thereof. The preservation No. of the virus strain is CGMCC NO.7606. The virus strain is duck-sourced H4N6 subtype avian influenza virus containing 8 fragments of which the CDS sequence shown in SEQ ID NO.1-SEQ ID NO.8, wherein the 8 fragments respectively correspond to PB2, PB1, PA, HA, NP, NA, M and NS. Whole genome sequencing is carried out on the virus strain, and the information is used for research on the detection, distribution and epidemic rule of the avian influenza virus as well as control of prevalence and spreading of the avian influenza. The virus strain supplements valuable genetic information of the H4N6 subtype avian influenza virus in China, provides valuable information for molecular evolution and epidemiologic study of H4N6 subtype avian influenza virus and can be used for preparing reagents for rapidly diagnosing H4N6 subtype influenza virus infection and vaccine study.

Description

technical field [0001] The invention belongs to the field of animal virology, and relates to a new isolated virus strain and its application, in particular to a duck-derived H4N6 subtype avian influenza virus strain and its application. Background technique [0002] Avian influenza (AI) is an infectious disease caused by type A influenza virus that seriously endangers the health of poultry. After poultry are infected with avian influenza virus (AIV), the symptoms can manifest as non-dominant infection, subclinical infection, or mild respiratory disease, decreased egg production to acute systemic fatal disease and other forms. According to its pathogenicity, AIV can be divided into highly pathogenic AIV (Highly Pathogenic Avian Influenza virus, HPAIV) and low pathogenic AIV (Low Pathogenic Avian Influenza virus, LPAIV). The H4 subtype is a low pathogenic AIV, which is mostly isolated from waterfowl and wild birds, persists in ducks, geese, etc., and may be transmitted to chi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00G01N33/569A61K39/145A61P31/16C12N15/44C12R1/93
Inventor 杨德全刘佩红周锦萍鞠厚斌葛菲菲刘健王建葛杰邓波
Owner SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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