Artificially recombined h5n8 influenza virus and its preparation method and application

A technology of influenza virus and avian influenza virus, which is applied in the direction of DNA preparation, recombinant DNA technology, and microbial-based methods, to achieve the effects of reducing production costs, improving vaccine quality, and good chicken embryo adaptability

Active Publication Date: 2022-06-14
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 h5n8 influenza virus and its preparation method and application
  • Artificially recombined h5n8 influenza virus and its preparation method and application
  • Artificially recombined h5n8 influenza virus and its preparation method and application

Examples

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Embodiment 1

[0039] Example 1: Preparation and Identification of Recombinant H5N8 Influenza Virus H5-Re14

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

[0041] 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 artificially recombined H5N8 influenza virus, a preparation method and application thereof. An artificial recombinant H5N8 influenza virus of the present invention has highly pathogenic avian influenza virus A / swan / Shanxi / 4‑1 / 2020 (H5N8) surface antigen hemagglutinin (HA) and neuraminidase (NA) genes , but the HA cleavage site shows the molecular characteristics of typical low pathogenic avian influenza virus (‑RETR‑), and has the PB2, PB1, PA, There are 6 internal genes including NP, M and NS, the strain number is H5‑Re14, and the deposit number is CCTCC NO: V202160. It has good chicken embryo adaptability, and the virus growth titer can be increased by more than 9 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 fields 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-sense 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 outbreaks 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 Patents(China)
IPC IPC(8): C12N7/01C12N7/04C12N15/44C12N15/56C12N15/85C12N15/10A61K39/145A61P31/16C12R1/93
CPCC12N7/00C07K14/005C12N9/2402C12N15/85C12N15/102A61K39/12A61P31/16C12Y302/01018C12N2760/16121C12N2760/16122C12N2760/16162C12N2800/107C12N2830/205C12N2760/16134A61K2039/552A61K2039/5254C12Q2531/113C12Q2521/107
Inventor 陈化兰曾显营田国彬李雁冰邓国华李呈军
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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