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Recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40/PR8) as well as preparation method and application of inactivated vaccine strain

An influenza virus, inactivated vaccine technology, applied in the direction of virus/phage, biochemical equipment and methods, antiviral agents, etc., can solve the problems of isolation, difficulty and high cost

Active Publication Date: 2014-12-24
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 strains with the above conditions from nature as vaccine strains
In China, due to the poor adaptability of chicken embryos isolated from wild strains, the current domestic avian-like H1N1 influenza vaccines have disadvantages such as high cost and short vaccine immunity period.

Method used

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  • Recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40/PR8) as well as preparation method and application of inactivated vaccine strain
  • Recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40/PR8) as well as preparation method and application of inactivated vaccine strain
  • Recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40/PR8) as well as preparation method and application of inactivated vaccine strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1: Screening, establishment of identification recombinant virus method

[0059] Utilize Trizol strong fission agent method, extract the RNA of influenza virus A / PR / 8 / 34 (H1N1) and avian type H1N1 influenza strain A / swine / Jiangsu / 40 / 2011 (H1N1), carry out reverse transcription, then use The fragment-specific identification primers of influenza virus A / PR / 8 / 34 (H1N1) and avian-like H1N1 influenza strain A / swine / Jiangsu / 40 / 2011 (H1N1) designed above were used for PCR reaction. The fragment-specific identification primers designed according to the gene of influenza virus A / PR / 8 / 34 (H1N1) can only amplify 8 specific fragments of influenza virus A / PR / 8 / 34 (H1N1) strain but cannot amplify Add any fragment of the avian H1N1 influenza isolate A / swine / Jiangsu / 40 / 2011(H1N1) strain, according to the genetic design of the avian H1N1 influenza isolate A / swine / Jiangsu / 40 / 2011(H1N1) The fragment-specific identification primers can only amplify 8 specific fragments of the av...

Embodiment 2

[0063] Example 2: Preparation and Identification of Recombinant Avian H1N1 Influenza Virus JS40 / PR8

[0064] 1. Co-infection - Recombination of two viral genomes:

[0065] 100 μl of avian-like H1N1 influenza virus domestic isolate strain A / swine / Jiangsu / 40 / 2011 (H1N1) and 50 μl of influenza virus A / PR / 8 / 34 (H1N1) (PR8 for short) were mixed and added to 850 μl of PBS for co-inoculation The allantoic cavity of 10-day-old SPF chicken embryo underwent natural reorganization. After culturing at 35°C for 24 hours, the allantoic fluid was collected.

[0066] 2. Antibody screening-removal of strains whose surface genes are derived from influenza virus A / PR / 8 / 34 (H1N1):

[0067] Take 50 μl of the allantoic fluid harvested under the above-mentioned specific artificial conditions and mix with 600 μl of anti-A / PR / 8 / 34 (H1N1) antiserum diluted by 4 times, react at room temperature for 30 minutes, and then inoculate each dilution Three chicken embryos. Next, the inoculated chicken embry...

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Abstract

The invention discloses a recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40 / PR8) as well as a preparation method and an application of the inactivated vaccine strain. The recombinant avian H1N1 influenza virus inactivated vaccine strain takes A / swine / Jiangsu / 40 / 2011 (H1N1) as genetic donors of surface hemagglutinin (HA) and neuraminidase (NA), and influenza virus A / PR / 8 / 34(H1N1) as donors of six internal genes of a polymerase PB2 subunit, a polymerase PB1 subunit, a polymerase PA subunit, nucleoprotein (NP), a matrix protein (M) and non-structural protein (NS); the vaccine strain is obtained by natural recombination; and the preservation number of the vaccine strain is CCTCC NO:V201419. The invention also discloses a method for building the recombinant homologous avian H1N1 influenza virus inactivated vaccine strain, and an application of the homologous recombinant avian H1N1 influenza virus inactivated vaccine strain for preventing homologous avian H1N1 influenza of animals or people.

Description

technical field [0001] The present invention relates to a recombinant genetic engineering vaccine, specifically, the present invention relates to a recombinant genetic engineering vaccine for preventing animal and human avian H1N1 influenza, more specifically, the vaccine involved in the present invention is a vaccine for preventing animal and human Inactivated recombinant avian H1N1 influenza virus vaccine for human avian H1N1 influenza. Background technique [0002] Influenza virus is an enveloped virus composed of segmented negative-sense RNA that can widely infect birds and mammals. In the world, influenza often occurs in the form of sporadic outbreaks and random pandemics. In the past few decades, the main choice of various countries is still the whole virus inactivated vaccine, which mainly stimulates the body to produce humoral immune resistance. Subtype of the hemagglutinin surface glycoprotein. [0003] Avian-like type H1N1 is widespread in pig herds in my country...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/145A61P31/16C12R1/93
Inventor 杨焕良陈化兰陈艳乔传玲
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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