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Anti-Newcastle disease, infectious bronchitis and bird flu vaccine composition and preparation

A vaccine composition and technology for bronchitis, applied in the direction of microorganisms, antiviral agents, virus antigen components, etc., can solve the problems of poor protection effect of H9 subtype avian influenza, risk of spreading poison, etc., achieve extensive cross-immunity, avoid spreading Toxic hazard, extensive cross-protective effect

Active Publication Date: 2016-12-28
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high variability of avian influenza, the current commercial vaccines have poor protection against H9 subtype avian influenza
At the same time, most of the H9 subtype vaccine antigens used in current commercial vaccines are inactivated wild virus, that is, whole virus antigens, and there is a risk of loose virus due to incomplete inactivation

Method used

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  • Anti-Newcastle disease, infectious bronchitis and bird flu vaccine composition and preparation
  • Anti-Newcastle disease, infectious bronchitis and bird flu vaccine composition and preparation
  • Anti-Newcastle disease, infectious bronchitis and bird flu vaccine composition and preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Isolation, Identification and Immunogenicity Analysis of SZ Strain of Avian Influenza Virus (H9 Subtype)

[0064] 1. Sample

[0065] It was isolated and identified by Pulaike Bioengineering Co., Ltd.

[0066] 2. Virus Isolation

[0067] The chicken swab samples collected from Shandong were centrifuged at 3000rpm for 5 minutes, and the dose of 0.2ml / embryo was aseptically inoculated into 10-day-old SPF (Specific pathogen Free, SPF) chicken embryos by the allantoic cavity route. . Continue incubation at 37°C. Eggs were illuminated every day, and chicken embryos that died within 24 hours were discarded, and all chicken embryos that died within 48 to 72 hours of incubation and survived within 72 hours were detected.

[0068] 3. Virus identification

[0069] (1) Red blood cell agglutination test

[0070] The HA titer of the harvested chicken embryo fluid was measured according to conventional methods, and the HA titer was 1:256.

[0071] (2) Virus subtype ide...

Embodiment 2

[0086] Embodiment 2: Construction of recombinant avian influenza virus (H9 subtype) SZ-1 strain

[0087] The invention provides a recombinant avian influenza virus (H9 subtype) SZ-1 strain, which is a 2:6 recombinant virus strain constructed by using the avian influenza virus (H9 subtype) SZ strain and artificial natural recombination technology. The gene comes from the donor strain avian influenza virus (H9 subtype) SZ strain, and the 6 internal genes come from the donor strain influenza virus PR8 (H1N1) strain.

[0088] 1. Co-infection - recombination of two viral genomes

[0089] Mix 100 μl of avian influenza virus (H9 subtype) SZ strain virus solution with 50 μl of influenza (H1N1 subtype) A / PR / 8 / 34 strain virus solution, add 850 μl of PBS (0.01mol / L, pH 7.0-7.4) 10-day-old SPF chicken embryos were inoculated through the allantoic cavity for natural recombination, cultured at 35°C for 24 hours, and the allantoic fluid was harvested.

[0090] 2. Antibody screening - remov...

Embodiment 3

[0097] Example 3: Identification of recombinant avian influenza virus (H9 subtype) SZ-1 strain

[0098] The recombinant avian influenza virus (H9 subtype) SZ-1 strain provided by the invention has extensive cross-immunity, high-titer chicken embryo adaptability, and no pathogenicity to chickens and chicken embryos.

[0099] 1. Determination of growth curve of recombinant virus

[0100] Inoculate five groups of SPF chicken embryos with the parental avian influenza virus (H9 subtype) SZ strain and the recombinant virus respectively, and culture them at 37°C for 24, 48, 72 and 96 hours respectively, and take the chicken embryos cultured for different times for hemagglutination Valence determination, detection of growth curves of parental avian influenza virus (H9 subtype) SZ strain and recombinant virus. see results figure 2 . The preparation of avian influenza vaccine strains requires not only good immunogenicity, but also good growth characteristics in production. The growt...

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Abstract

The invention relates to a recombinant avian influenza virus strain. The recombinant avian influenza virus strain is obtained by recombining influenza virus strains and avian influenza virus strains through genetic recombination. It has no pathogenicity to chickens and chicken embryos, is safer than wild strains, and can effectively avoid inactivation Divergence hazard caused by incompleteness. The invention also relates to a vaccine composition against chicken Newcastle disease, infectious bronchitis and H9 subtype bird flu containing the recombinant bird flu virus strain. The vaccine composition can not only prevent Newcastle disease and infectious bronchitis, but also has good cross immunity to most H9N2 subtype avian influenza viruses currently prevalent in my country. Therefore, the vaccine composition has broad cross-protection against H9N2 subtype avian influenza compared with current commercialized vaccines.

Description

technical field [0001] The invention belongs to the technical field of animal husbandry biopharmaceuticals, and relates to a recombinant avian influenza virus strain, in particular to a recombinant avian influenza virus strain and an anti-Newcastle disease, infectious bronchitis and avian influenza vaccine composition containing the strain. Background technique [0002] Avian influenza (AI) is an infection and / or disease syndrome in poultry and other birds caused by influenza A virus of the Orthomyxoviridae family. After infection, housed birds (including poultry) may show clinical symptoms, respiratory Systemic diseases, decreased egg production, and acute hemorrhagic diseases with a high fatality rate can also manifest as subclinical symptoms, mild respiratory infections, and asymptomatic infection. Among them, the H9N2 subtype avian influenza virus has been widely distributed and prevalent in my country since my country first reported the isolation of the H9N2 subtype avi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01A61K39/145A61K39/215A61K39/17A61P31/16A61P31/14
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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