Artificially recombined h5n6 influenza virus and its preparation method and application

A technology for 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 virus isolation and difficulty, and achieve the effects of reducing production cost, high biological safety, and improving vaccine quality.

Active Publication Date: 2022-06-21
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 viruses with the above conditions from nature as vaccine strains

Method used

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  • Artificially recombined h5n6 influenza virus and its preparation method and application
  • Artificially recombined h5n6 influenza virus and its preparation method and application
  • Artificially recombined h5n6 influenza virus and its preparation method and application

Examples

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Embodiment 1

[0039] Example 1: Preparation and Identification of Recombinant H5N6 Influenza Virus H5-Re13

[0040] 1. Construction and identification of 2 surface genes (HA and NA) of the recombinant virus:

[0041] RNA of H5N6 subtype highly pathogenic avian influenza isolate DK / FJ / S1424 / 20 (H5N6) was extracted and reverse transcribed into cDNA using HA1 fragment-specific primers of the HA gene (HA1-U: atg gag aaa ata gta ctt ctt cttt; HA1-L: gct cca aac agt cct cta gtt tcc ctt aga gga cta ttt ctg agc, wherein HA1-L is a primer that mutates the HA cleavage site as a molecular feature of low pathogenicity virus) and HA2 Fragment-specific primers (HA2-U: actaga gga ctg ttt gga gct ata gca gga t; HA2-L: tta aat gca aat tct gca ttg taac) were used to amplify HA1 and HA2 of the HA gene by RT-PCR Fragment ( figure 1 Middle A), and while amplifying the HA1 gene fragment, carry out weakening mutation modification on the HA cleavage site; then use HA1 and HA2 as templates, HA1-U / HA2-L as primers...

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Abstract

The invention discloses an artificially recombined H5N6 influenza virus, a preparation method and application thereof. A kind of artificial recombinant H5N6 influenza virus of the present invention has highly pathogenic avian influenza virus A / duck / Fujian / S1424 / 2020 (H5N6) surface antigen hemagglutinin (HA) and neuraminidase (NA) gene, but The HA cleavage site shows the molecular characteristics of typical low pathogenic avian influenza virus (‑RETR‑), and has the PB2, PB1, PA, NP, There are 6 internal genes in M ​​and NS, the strain number is H5‑Re13, and the deposit number is CCTCC NO: V202159. It has good chicken embryo adaptability, and the virus growth titer can be increased by more than 10 times compared with wild virus; using this as a vaccine strain to produce vaccines can greatly reduce vaccine production costs and improve vaccine quality.

Description

technical field [0001] The invention relates to the field of reverse genetics technology and animal infectious disease technology, in particular to an artificially recombined and attenuated H5N6 subtype recombinant influenza virus and its preparation method and application. Background technique [0002] Highly pathogenic avian influenza (avian influenza, AI) is a severe respiratory disease caused by type A influenza virus, and is classified as a class A infectious disease by the World Organization for Animal Health (OIE). Economic loss, and a serious threat to human life, AI prevention and control has become the focus of research on zoonotic infectious diseases. Avian influenza virus (AIV) is a segmented negative-strand RNA virus containing 8 gene segments, which are prone to gene mutation and recombination to produce new viruses. According to the surface proteins of the virus, AIV is divided into 16 hemagglutinin (HA) and 9 neuraminidase (NA) subtypes. Various H5Nx subtyp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N15/56C12N15/44C12N15/85A61K39/145A61P31/16C12R1/93
CPCC12N7/00C12N9/2402C12N15/85C07K14/005A61K39/12A61P31/16C12Y302/01018C12N2760/16121C12N2760/16122C12N2760/16134A61K2039/5254A61K2039/552
Inventor 陈化兰曾显营田国彬邓国华施建忠李呈军
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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