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Novel antigen group H9N2 avian influenza virus and application thereof

A bird flu virus and antibody technology, applied in antiviral agents, antiviral immunoglobulin, virus/bacteriophage, etc., can solve the problem of inability to prevent bird flu virus

Inactive Publication Date: 2016-03-09
SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the above-mentioned technical problems in the prior art, the present invention provides a H9N2 avian influenza virus of a novel antigenic group and its use, and the H9N2 avian influenza virus of the novel antigenic group and its use solve the problems in the prior art. The inability to prevent new avian influenza viruses and lead to technical problems in the spread of new avian influenza outbreaks in poultry or humans

Method used

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  • Novel antigen group H9N2 avian influenza virus and application thereof
  • Novel antigen group H9N2 avian influenza virus and application thereof
  • Novel antigen group H9N2 avian influenza virus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The acquisition of embodiment 1 virus

[0022] Obtained from chickens in live poultry wholesale markets in Shanghai in 2013, collect live poultry samples including tracheal or cloacal swabs, and place the collected samples in isotonic phosphate buffered saline (PBS) with a pH value of 7.0-7.4 containing antibiotics . 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-11-day-old SPF chicken embryos through the allantoic cavity at an amount of 0.2mL / embryo, inoculate 5 embryos for each sample, and incubate in an incubator at 35°C-37°C. After 18 hours, The death of chicken embryos was observed every 8h. Aseptically collect dead embryos after 18 hours and chicken embryo allantoic fluid of surviving chicken embryos after 96 hours, measure hemagglutination activity, carry out gene...

Embodiment 2H9

[0023] The whole gene sequencing of embodiment 2H9N2 avian influenza virus

[0024] 2.1 Obtained from chickens in the live poultry wholesale market in Shanghai in 2013, a strain of H9N2 avian influenza virus was isolated and identified: A / chicken / Shanghai / 1107 / 2013 (preservation number: CGMCCNO: 10925). Firstly, the chicken trachea and cloaca samples were tested with H9 fluorescent RT-PCR kit, the H9 positive samples were treated and inoculated into the allantoic cavity of chicken embryos, and the isolated poisonous allantoic fluid was tested for HA by the β-micro method to detect its blood. coagulation and HA potency. According to the HA titer measured in the HA test, the isolated poisonous allantoic fluid was prepared into a solution of 4 units, and the HI test was carried out with Newcastle disease positive serum, avian influenza H5 and H9 positive serum to detect the blood of the isolate. Whether coagulation can be inhibited by avian influenza H9 positive serum, the detai...

Embodiment 3

[0075] Example 3 virus strain and gene sequence number

[0076] 1) A / chicken / Shanghai / Y1 / 2002 (GQ335463-GQ335470),

[0077] 2) A / chicken / Shanghai / Y1 / 2006 (GQ335471-GQ335478),

[0078] 3) A / duck / Shanghai / C60 / 2007 (KC768039-KC768046),

[0079] 4) A / chicken / Shanghai / Y1 / 2007 (GQ335487-GQ335494),

[0080] 5) A / chicken / Shanghai / Y2 / 2008 (GQ335511-GQ335518),

[0081] 6) A / duck / Shanghai / C163 / 2009 (KC768047-KC768054),

[0082] 7) A / swine / Henan / Y1 / 2009 (KC768047, KC768049, KC768051, KC768053, KC768055, KC768057, KC768059, KC768061),

[0083] 8) A / chicken / Shanghai / A / 2010 (KJ726689, KJ726695, KJ726701, KJ7267007, KJ726713, KJ726719, KJ726725, KJ726731),

[0084] 9) A / chicken / Shanghai / C / 2011 (KJ726691, KJ726697, KJ726703, KJ726709, KJ726715, KJ726721, KJ726727, KJ726733),

[0085] 10) A / chicken / Shanghai / C1 / 2012 (KC417046, KC417049, KC417052, KC417055, KC417058, KC417061, KC417064, KC417067),

[0086] 11) A / chicken / Shanghai / C2 / 2012 (KC417047, KC417050, KC417053, KC417056, KC417059, KC...

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Abstract

The invention provides a separate H9N2 avian influenza virus. A genome of the H9N2 avian influenza virus is composed of RNA fragments shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8. The invention further provides a vaccine used for preventing the H9N2 avian influenza virus and a preparation method of the vaccine. The preparation method includes the steps that vaccine antigen liquid is prepared, and the toxic value is measured; the antigen liquid is inactivated; oil phase adjuvant is prepared; a vaccine aqueous phase is prepared; the vaccine is prepared. By inoculating the vaccine, infection of the antigen group D H9N2 avian influenza virus can be prevented. By using an antibody, infection of the antigen group D H9N2 avian influenza virus can be treated.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to an avian influenza virus, in particular to a novel antigen group H9N2 avian influenza virus and its application. Background technique [0002] H9N2 subtype avian influenza virus (Avian influenza virus, AIV) has been widely prevalent in my country and around the world since the 1990s, not only causing huge economic losses to the poultry industry, but also immeasurable hidden dangers to public health safety. It also poses a huge threat to human health. In my country, the unique geographical environment and breeding mode provide favorable conditions for the occurrence and spread of this disease. The immune pressure and the existence of the live poultry market provide the necessary conditions for the recombination and mutation of the H9N2 subtype avian influenza virus. At present, the H9N2 subtype vaccine has been used for routine immunization, and the outbreak of H9N2 subtype AIV has been...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/04A61K39/145A61P31/16C07K16/10G01N33/569
Inventor 葛菲菲刘健李鑫鞠厚斌齐新永杨德全徐锋周锦萍
Owner SHANGHAI ANIMAL EPIDEMIC PREVENTION & CONTROL CENT