Avian influenza virus (AIV) as well as kit and vaccine used for detecting and preventing same

An avian influenza virus and kit technology, applied in the direction of antiviral agents, antiviral immunoglobulins, viruses/phages, etc., can solve the problems of inability to identify and prevent avian influenza viruses, transmission, etc.

Inactive Publication Date: 2013-01-02
SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention provides three kinds of H9N2 avian influenza viruses and kits and vaccines for detecting and preventing avian influenza viruses. Solve the technical problem that new avian influenza viruses cannot be confirmed and prevented in the existing technology, which leads to the spread of new avian influenza epidemics in birds or humans

Method used

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  • Avian influenza virus (AIV) as well as kit and vaccine used for detecting and preventing same
  • Avian influenza virus (AIV) as well as kit and vaccine used for detecting and preventing same
  • Avian influenza virus (AIV) as well as kit and vaccine used for detecting and preventing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] The acquisition of embodiment 1 virus

[0100]From 2002 to 2008, commercial Dai Sanhuang chickens were obtained from live poultry wholesale markets in Shanghai. Live poultry samples were collected, including tracheal or cloacal swabs, and the collected samples were placed in isotonic phosphate buffer solution with a pH value of 7.0 to 7.4 containing antibiotics. (PBS). Fully twist and wring out the cotton swab identified as a positive sample, discard the swab, centrifuge the sample solution at 1000r / min for 10min, and take the supernatant as the inoculation material. Take the processed sample and inoculate 9-day-old to 11-day-old SPF chicken embryos through the allantoic cavity at an amount of 0.2 mL / embryo, inoculate 5 embryos for each sample, and incubate in an incubator at 35°C to 37°C. After 18 hours, The death of chicken embryos was observed every 8h. Aseptically collect the dead embryos after 18 hours and the chick embryo allantoic fluid of the surviving chicken...

Embodiment 2

[0101] Example 2 Whole-gene sequencing of three genotypes of H9N2 avian influenza virus

[0102] 2.1 From 2002 to 2008, commercial Dai Sanhuang chickens were obtained from live poultry wholesale markets in Shanghai, and three genotypes of H9N2 avian influenza virus were isolated and identified: A / Chicken / Shanghai / Y1 / 06 (abbreviated as Ck / SH / Y1 / 06, genotype Y1); A / Chicken / Shanghai / Y1 / 07 (abbreviated as Ck / SH / Y1 / 07, genotype Y2); A / Chicken / Shanghai / Y1 / 08 (abbreviated as Ck / SH / Y1 / 08, genotype Y3). The H9 subtype serum used for virus identification was purchased from Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The specific primers designed to amplify all 8 gene fragments of H9 subtype AIV were synthesized by Treasure Biological (Dalian) Engineering Company.

[0103] HA gene upstream primer P1: 5'-CAGGGGAATTTCACAACCAGTCAAA-3';

[0104] Downstream primer P2: 5'-GCCAGGGTGTTTTTGCCAATTATAC-3'.

[0105] M gene upstream primer P1: 5'-AAGATGAGCCTT...

Embodiment 3

[0170] Example 3 HA Gene Amino Acid Sequence Analysis

[0171] The deduced amino acid sequence analysis showed that the three genotype strains all contained 7 potential glycosylation sites (Asn-X-Ser / Thr, X is any amino acid except Pro), 5 of which were located in the HA1 part ( 29aa~31aa, 141aa~143aa, 218aa~220aa, 298aa~300aa, 305aa~307aa), and 2 are located in the HA2 part (492aa~494aa, 551aa~553aa). Sequence analysis of the cleavage site revealed that all the strains were RSSR↓GLF and had low pathogenicity to poultry. The amino acids (109aa, 161aa, 163aa, 191aa, 202aa, 203aa) constituting the receptor binding site were relatively conserved in all strains except for the 198th amino acid with a large change. In addition, this study shows that Ck / SH / Y1 / 06 is Gln at site 226 of the HA gene, which proves to be human non-susceptible H9 subtype AIV, while the other two strains are Leu at this site, indicating that they are capable of infecting humans. tropism (as shown in Table ...

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Abstract

The invention relates to an avian influenza virus (AIV) as well as a kit and a vaccine used for detecting and preventing the same, belonging to the technical field of biology, relating to a novel strain of separated AIV and disclosing H9N2 AIVs of three types, a kit for detecting H9N2 AIVs of three types, a vaccine for preventing H9N2 AIVs of three types and an antibody against H9N2 AIV. By utilizing the kit of the invention, the possibility whether the avian influenza is caused by the H9N2 AIVs of three types in the invention can be rapidly detected or removed, thereby controlling the epidemic situation of the avian influenza as soon as possible; and simultaneously, by inoculating the vaccine of the invention, infection due to the H9N2 AIVs of three gene types can be prevented. The invention also can provide biological information for an AIV monitoring system.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and relates to three new strains of H9N2 avian influenza virus, a kit and a vaccine for detecting and preventing the avian influenza virus. Background technique: [0002] Avian influenza (AI) is an infectious disease caused by type A influenza virus that seriously endangers the health of poultry. After poultry are infected with avian influenza virus (AIV), the symptoms can manifest as non-dominant infection, subclinical infection, or mild respiratory disease, decreased egg production to acute systemic fatal disease and other forms. According to the difference in pathogenicity of AIV to susceptible chickens, AIV can be divided into highly pathogenic AIV and low pathogenic AIV. The avian influenza caused by highly pathogenic AIV (HPAIV) infection of poultry has a devastating blow to the poultry industry, so it is stipulated as a class A disease by the International Office of Epizootics; and some low-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/68C07K16/10A61K39/145A61P31/16C12R1/93
Inventor 刘佩红周锦萍葛菲菲刘健张维谊鞠厚斌李凯航
Owner SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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