Artificially recombined h7n9 influenza virus and its preparation method and application

A technology of influenza virus and avian influenza virus, which is applied in the field of reverse genetics technology and animal infectious diseases, can solve the problems of isolating influenza virus strains, difficulty, etc., and achieve the goals of reducing production costs, high biological safety, and improving vaccine quality Effect

Active Publication Date: 2022-06-07
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to isolate influenza strains with the above conditions from nature as vaccine strains

Method used

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  • Artificially recombined h7n9 influenza virus and its preparation method and application
  • Artificially recombined h7n9 influenza virus and its preparation method and application
  • Artificially recombined h7n9 influenza virus and its preparation method and application

Examples

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Embodiment 1

[0039] Example 1: Preparation and identification of recombinant H7N9 influenza virus H7-Re4

[0040] 1. Construction and identification of two surface genes (HA and NA) of recombinant virus:

[0041] Using the viral RNA extraction kit, the RNA of the highly pathogenic avian influenza virus isolate CK / YN / SD024 / 21 (H7N9) of H7N9 subtype in my country was extracted as a template, and the HA1 fragment-specific primer (HA1-U: atg aac act caa) was used as a template. atc ctgata ttc g; HA1-L-: gca cca aat agg cct cgt ccc ttt gga acc tca gga aca ttc tt, where HA1-L is a primer that simultaneously mutates the HA cleavage site for low pathogenicity molecular characteristics) and HA2 fragment Specific primers (HA2-U: gga cga ggc cta ttt ggt gct ata gcg g; HA2-L: tta tat aca aat agt gcaccg cat g) were used to amplify the HA1 and HA2 fragments of HA respectively by RT-PCR ; Then take HA1 and HA2 as templates, HA1-U / HA2-L as primers, use phusion high-fidelity polymerase to carry out overlap e...

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Abstract

The invention discloses an artificially recombined H7N9 influenza virus, a preparation method and application thereof. The artificial recombinant H7N9 influenza virus of the present invention has highly pathogenic avian influenza virus A / chicken / Yunnan / SD024 / 2021 (H7N9) surface antigen hemagglutinin (HA) and neuraminidase (NA) gene, but HA cracks The amino acid sequence of the site is ‑PKGR‑, with 6 internal genes including PB2, PB1, PA, NP, M and NS of influenza virus A / PR / 8 / 34 (H1N1), the strain number is H7‑Re4, and the deposit number is CCTCC NO: V202161. The virus strain has good adaptability to chicken embryos, and the virus growth titer can be increased by more than 9 times compared with the wild virus; it can be used as a vaccine strain to produce vaccines, which can greatly reduce the production cost of vaccines and improve the quality of vaccines.

Description

technical field [0001] The invention relates to the technical field of reverse genetics technology and animal infectious diseases, in particular to an artificially recombined and weakened H7N9 subtype recombinant influenza virus and a preparation method and application thereof. Background technique [0002] Avian influenza virus (AIV) is a segmented negative-strand RNA virus that can broadly infect birds and mammals. AIV is classified into 16 hemagglutinin (HA) and 9 neuraminidase (NA) subtypes according to the surface proteins of the virus. Among them, H5 and H7 subtypes of AIV can cause outbreaks in poultry or wild birds, resulting in significant economic losses, and can also infect humans and cause public health hazards. China has developed a series of inactivated whole virus vaccines, which have successfully controlled H5 subtype since 2004 and H7N9 subtype avian influenza since 2017. [0003] In order to deal with the poultry epidemic situation and public health hazar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N15/44C12N15/85A61K39/145A61P31/16C12R1/93
CPCC12N7/00C07K14/005C12N15/85A61K39/12A61P31/16C12N2760/16121C12N2760/16122C12N2760/16151C12N2760/16134C12N2800/107
Inventor 陈化兰曾显营田国彬施建忠邓国华李呈军
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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