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Avian influenza H9 subtype inactivated vaccine, preparation method and application thereof

An inactivated vaccine, avian influenza technology, applied in biochemical equipment and methods, microorganism-based methods, pharmaceutical formulations, etc. Drop and other problems, to achieve the effect of good attack protection, stable efficacy and strong immunogenicity

Active Publication Date: 2015-07-15
ZHAOQING DAHUANONG BIOLOGIC PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the widespread application of single or combined vaccines of H9 subtype avian influenza vaccines, in recent years, under the effect of immune pressure, the immune protection of previously used vaccine strains against field isolated strains began to decline. According to the needs of self-immunity, many breeding units have used inactivated vaccines made of local circulating strains to prevent and control the occurrence of H9 subtype avian influenza, but these vaccines cannot be formally produced, sold and widely promoted, and their safety cannot be guaranteed

Method used

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  • Avian influenza H9 subtype inactivated vaccine, preparation method and application thereof
  • Avian influenza H9 subtype inactivated vaccine, preparation method and application thereof
  • Avian influenza H9 subtype inactivated vaccine, preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0024] Embodiment 1: Research on the biological characteristics of H9 subtype avian influenza virus HZ strain and FJ strain

[0025] The H9 subtype avian influenza virus HZ strain and FJ strain described in the present invention were isolated from diseased materials collected from diseased chicken flocks in the Huzhou area of ​​Zhejiang Province and Putian area of ​​Fujian Province in 2010 respectively, and were identified by sequencing. H9 subtype of avian influenza virus. Its biological characteristics are studied as follows: 1) Virus seed passage: the primary seed virus (E1 generation) of HZ strain and FJ strain was diluted to 10 with sterilized normal saline. -4 , inoculate 9-day-old SPF chicken embryos into the allantoic cavity, 0.2 mL per embryo, continue to incubate at 37°C for 96 hours, harvest dead and surviving chicken embryos within 24 to 96 hours, harvest chicken embryo liquid one by one after freezing overnight Detect its HA titer, mix the chicken embryo fluid te...

Embodiment 2

[0053] Embodiment 2: Safety research of avian influenza H9 subtype inactivated vaccine

[0054] A kind of preparation method of avian influenza H9 subtype inactivated vaccine is characterized in that comprising the steps:

[0055] 1) Preparation of H9 subtype avian influenza virus HZ strain venom: make H9 subtype avian influenza virus HZ strain with normal saline for 10 -4 Dilute, inoculate 9-11 day-old SPF chicken embryos, aseptically harvest chicken embryo allantoic fluid with HA≥8log2 that died within 24-96 hours and survived after 96 hours as H9 subtype avian influenza virus HZ strain venom;

[0056] 2) Preparation of H9 subtype avian influenza virus FJ strain venom: make H9 subtype avian influenza virus FJ strain with normal saline for 10 -4 Dilute, inoculate 9-11 day-old SPF chicken embryos, aseptically harvest chicken embryo allantoic fluid with HA≥8log2 that died within 24-96 hours and survived after 96 hours as H9 subtype avian influenza virus FJ strain venom;

[00...

Embodiment 3

[0068] Embodiment 3: Avian influenza H9 subtype inactivated vaccine is to SPF chicken immune effect test

[0069] Prepare avian influenza H9 subtype inactivated vaccine according to the method of Example 2, immunize 21-day-old SPF chickens with the prepared avian influenza H9 subtype inactivated vaccine, 0.3mL / only, take blood 21 days after immunization, separate serum, measure HI Antibody titer, at the same time, the subwing veins were used to attack the isolated strains of AIV H9 subtypes in different places, and the challenge dose was 0.2mL / bird. On the 5th day after the challenge, the throat swabs of each chicken were collected and inoculated through the allantoic cavity Viruses were isolated from SPF chicken embryos aged 9-10 days. The throat swab sample of each chicken was inoculated into the allantoic cavity of 5 SPF chicken embryos aged 9-10 days, and incubated for 96 hours. Regardless of the dead embryos or live embryos, the erythrocyte agglutination value of chicken ...

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Abstract

The present invention relates to an avian influenza H9 subtype inactivated vaccine, which is formed by mixing two strains of inactivated H9 subtype avian influenza viruses and an oil adjuvant, wherein the two strains of the inactivated H9 subtype avian influenza viruses are respectively the HZ strain and the FJ strain, the HA gene sequence of the HZ strain is represented by SEQ ID NO:1, and the HA gene sequence of the FJ strain is represented by SEQ ID NO:2. According to the preparation method, the virus solution of the HZ strain and the virus solution of the FJ strain of the H9 subtype avian influenza viruses are prepared and inactivated, the oil phase solution and the water phase solution are prepared, and emulsification is performed to obtain the finished product. According to the present invention, the two strains of the H9 subtype avian influenza viruses separated from different places are utilized to prepare the inactivated vaccine with characteristics of strong immunogenicity and good cross-protection property, wherein the inactivated vaccine can be used for prevention of chicken H9 subtype avian influenza diseases. In addition, after the inactivated vaccine is adopted to immunize chicken, the antibody titer is high so as to make the chicken have good virus challenge protection property on the H9 subtype strains epidemic in different places, the safety is high, and the efficacy is stable.

Description

technical field [0001] The invention belongs to the technical field of biological products for animals, and in particular relates to an inactivated avian influenza H9 subtype vaccine and a preparation method and application thereof. Background technique [0002] Avian Influenza (AI) is a general term for poultry infections and diseases caused by type A influenza viruses of the genus Influenzavirus. Poultry such as chickens, turkeys, ducks, and quails, as well as wild birds, waterfowl, and seabirds can be infected, and the harm caused to domestic chickens and turkeys is the most serious. The main reason for the worldwide epidemic of influenza, pigs can also be used as "mixers" to spread the disease. [0003] According to the difference in pathogenicity of avian influenza virus AIV to birds, AIV can be divided into highly pathogenic avian influenza virus (HPAIV) and low pathogenic avian influenza virus (LPAIV). H9 subtype avian influenza belongs to low pathogenicity influenz...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/145A61P31/16C12R1/93
Inventor 陈瑞爱林绮萍陈春丽黄文科徐家华严洁珍
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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