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H9 subtype avian influenza virus isolate and application thereof

An avian influenza virus and avian influenza technology, applied in the field of animal virology, can solve the problems of HA and NA antigenic changes, the decline of immune protection efficacy of vaccine strains, and immune failure, etc., and achieve good immune prevention, good cross-immunogenicity, Good effect on immunogenicity

Active Publication Date: 2021-03-30
PU LIKE BIO ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When this subtype of virus simply infects chickens, it has a high morbidity rate and low mortality rate, and only mild respiratory symptoms appear, causing a decrease in egg production rate and hatching rate. Caused a high lethality rate of poultry, so it is very harmful to the poultry industry
[0004] Vaccination is currently one of the main means of preventing and controlling H9N2 subtype avian influenza, but the virus can produce mutant strains through antigenic drift and antigenic transformation, resulting in changes in the antigenicity of HA and NA, resulting in immune failure
Especially in recent years, poultry flocks have been infected with H9N2 subtype avian influenza, even in immune populations. The reason for this phenomenon is that there is a large genetic distance and antigenic Sexual differences, leading to a decrease in the immune protection efficacy of vaccine strains

Method used

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  • H9 subtype avian influenza virus isolate and application thereof
  • H9 subtype avian influenza virus isolate and application thereof
  • H9 subtype avian influenza virus isolate and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Isolation and identification of embodiment 1 virus

[0066] The low-pathogenic H9N2 subtype avian influenza virus has spread widely in my country. In recent years, chickens immunized with H9N2 subtype avian influenza vaccines have been infected with H9N2 subtype avian influenza and died, suggesting that the virus may drift through antigen Or the new epidemic trend of antigen transformation and evolution, which eventually leads to immune failure. Therefore, a large number of epidemiological investigations and researches were carried out on the mutation of the virus, and finally a strain of H9 was successfully isolated from a chicken farm in Hefei City, Anhui Province. Subtype of avian influenza virus.

[0067] 1.1 Isolation of virus

[0068] Select the diseased chickens with symptoms of respiratory system such as coughing and sneezing, bronchial embolism, tracheal mucosal edema, and pathological changes such as serous exudate, and get their internal organs (liver, kidney,...

Embodiment 2

[0089] Embodiment 2 is measured to chicken embryo and chicken pathogenicity

[0090] 2.1 Determination of half infection dose (EID50) of chicken embryo

[0091] The isolated virus was serially diluted 10 times with sterilized physiological saline, and 10 -1 ~10 -9 Inoculate 10-11-day-old SPF chicken embryos through the allantoic cavity with diluted virus liquid, 0.1ml / piece, inoculate 5 chicken embryos for each dilution, discard 24-hour dead embryos, and freeze overnight at 4°C 120 hours after inoculation For dead chicken embryos, the HA titer was measured embryo by embryo. If the HA titer was greater than 1:16, it was determined to be positive, and the virus EID was calculated by referring to the Reed-Muench method. 50 for 10 9.17 / 0.1ml.

[0092] 2.2 Toxicity to SPF chicken embryos

[0093] Dilute the HF strain E1 generation virus solution with sterilized physiological saline for 10 5 times, inoculate 10 10-day-old SPF chicken embryos through the allantoic cavity, 0.1m...

Embodiment 3

[0096] Example 3 Analysis of virus antigenicity

[0097] 3. Preparation of 1H9 single-factor serum

[0098] The 10 isolated strains of H9N2 subtype AIV and the HF strain involved in the present invention were used to prepare oil emulsion inactivated vaccines respectively. Immunize 21-day-old SPF chickens with the above-mentioned vaccines, 3 chickens for each vaccine, and carry out secondary immunization 14 days after the first immunization. Each time, 0.3ml of the vaccine is immunized through the leg muscles, and they are raised in a positive pressure isolator for poultry. 21 days after the second immunization, blood was collected aseptically, and the serum was separated to prepare single-factor positive serum, which was packed into small tubes and stored at -20°C.

[0099] 3.2 Determination of antigenic cross-reactivity between different strains

[0100] The 11 strains of H9N2 subtype AIV prepared in 3.1 were used to configure 4 units of antigen respectively, and then the h...

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Abstract

The invention provides an H9 subtype avian influenza HF strain and a vaccine composition prepared from the inactivated antigen of the H9 subtype avian influenza HF strain. The H9 subtype avian influenza HF strain has good immunogenicity, the existing H9 subtype avian influenza can be completely protected under the condition of low content, and the H9 subtype avian influenza HF strain can be used for preparing combined vaccines and compound vaccines together with various other antigens.

Description

technical field [0001] The invention relates to the field of animal virology, in particular to a method for isolating, identifying and purifying an H9N2 avian influenza virus strain, and an inactivated vaccine for preventing avian influenza prepared by using the strain. Background technique [0002] Avian influenza (Avian influenza, AI) is an acute, febrile, highly contagious avian infectious disease caused by avian influenza virus (AIV), which is a single-stranded negative-sense segmented RNA virus. Belongs to Orthomyxoviridae, Influenza A virus genus. According to the different pathogenicity and virulence of the virus, it can be divided into highly pathogenic avian influenza virus and low pathogenic avian influenza virus clinically, and according to its surface glycoprotein hemagglutinin (Hemagglutinin, HA) and neuraminic acid The difference of the enzyme (Neuraminidase, NA) can divide the type A influenza virus into different subtypes. Currently, there are 16 HA subtypes...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/145A61K39/235A61K39/17A61K39/215A61K39/12A61K39/39A61P31/14A61P31/16A61P31/20C12R1/93
CPCC12N7/00A61K39/12A61K39/39A61P31/16A61P31/14A61P31/20C12N2760/16121C12N2760/16134C12N2710/10221C12N2710/10234C12N2760/18134C12N2770/20034C12N2720/10034A61K2039/5252A61K2039/552A61K2039/55566A61K2039/70Y02A50/30
Inventor 田克恭王婉冰张许科
Owner PU LIKE BIO ENG
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