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Artificial recombinant H5N8 influenza virus and preparation method and application thereof

An influenza virus, avian influenza virus technology, applied in the field of reverse genetics technology and animal infectious diseases, can solve the problem of isolating influenza strains, difficult and other problems, achieve good chicken embryo adaptability, reduce production costs, and improve vaccine quality Effect

Active Publication Date: 2022-01-11
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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  • Artificial recombinant H5N8 influenza virus and preparation method and application thereof
  • Artificial recombinant H5N8 influenza virus and preparation method and application thereof
  • Artificial recombinant H5N8 influenza virus and preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0040] Example 1: Preparation and Identification of Recombinant H5N8 Influenza Virus H5-Re14

[0041] 1. Construction and identification of 2 surface genes (HA and NA) of the recombinant virus:

[0042] The RNA of H5N8 subtype highly pathogenic avian influenza isolate SW / SX / 4-1 / 20 (H5N8) was extracted and reverse-transcribed into cDNA using HA1 fragment-specific primers of the HA gene (HA1-U: atg gag aac ata gta ctt ctt ctt g; HA1-L-: gcc ccg aac agg cct cta gtt tct ctt aga gga cta ttt ctg agc, where HA1-L is a mutant HA cleavage site with low pathogenicity virus molecular characteristics Primers) and HA2 fragment-specific primers (HA2-U: actaga ggc ctg ttc ggg gcg ata gca ggg t; HA2-L: tta aat gca aat tct gca ctg taac), respectively amplified the HA gene by RT-PCR HA1 and HA2 fragments ( 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 artificial recombinant H5N8 influenza virus and a preparation method and application thereof. The artificial recombinant H5N8 influenza virus has a highly pathogenic avian influenza virus A / swan / Shanxi / 4-1 / 2020 (H5N8) surface antigen Hemagglutinin (HA) gene and a Neuraminidase (NA) gene, but an HA cleavage site has a typical low pathogenic avian influenza virus molecular characteristic (-RETR-), and has six internal genes of PB2, PB1, PA, NP, M and NS of an influenza virus chick embryo high-titer adaptive strain A / PR / 8 / 34(H1N1), the strain number is H5-Re14, and the preservation number is CCTCC NO: V202160. The artificial recombinant H5N8 influenza virus has good chick embryo adaptability, and the virus growth titer can be increased by 9 times or above compared with that of a wild virus; and the artificial recombinant H5N8 influenza virus is used as a vaccine strain for producing a vaccine, the production cost of the vaccine can be greatly reduced, and the quality of the vaccine is improved.

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 H5N8 subtype recombinant influenza virus and its preparation method and application. Background technique [0002] Avian influenza virus (AIV) is a segmented negative-strand RNA virus that can widely infect birds and mammals. According to the surface proteins of the virus, AIV is divided into 16 hemagglutinin (HA) and 9 neuraminidase (NA) subtypes. Among them, H5 and H7 subtypes of AIV can cause epidemics in poultry or wild birds, resulting in significant economic losses, and can also infect humans, causing public health hazards. China has developed a series of whole-virus inactivated vaccines, which have successfully controlled H5N1 since 2004 and H7N9 subtype avian influenza since 2017. [0003] In order to cope with the poultry H5N8 epidemic and public health hazards that may occur i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85C12N15/44C12N15/11A61K39/145A61P31/16C12R1/93
CPCC12N7/00C07K14/005C12N9/2402C12N15/85C12N15/102A61K39/12A61P31/16C12Y302/01018C12N2760/16121C12N2760/16122C12N2760/16162C12N2800/107C12N2830/205C12N2760/16134A61K2039/552A61K2039/5254C12Q2531/113C12Q2521/107
Inventor 陈化兰曾显营李雁冰邓国华田国彬李呈军
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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