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H9 subtype lowly pathogenic avian influenza virus and application thereof

A bird flu virus, low pathogenicity technology, applied in the direction of antiviral agent, virus/phage, medical preparations containing active ingredients, etc., can solve the problems such as the decline of vaccine protection effect, and achieve the prevention of H9 subtype bird flu, good Effect of specificity and good immunogenicity

Active Publication Date: 2018-01-09
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as the H9 subtype of avian influenza virus continued to mutate, the protective effect of earlier vaccines began to decline

Method used

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  • H9 subtype lowly pathogenic avian influenza virus and application thereof
  • H9 subtype lowly pathogenic avian influenza virus and application thereof
  • H9 subtype lowly pathogenic avian influenza virus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation and Identification of H9 Subtype Avian Influenza Virus X-13 Strain

[0030] 1. Epidemiological investigation

[0031] Investigate the clinical manifestations, necropsy lesions and mortality of chicken flocks. In December 2010, a flock of Sanhuang broiler chickens in a large chicken farm in Xinzheng, Henan Province began to suffer from symptoms such as loss of appetite, depression, coughing, head shaking, sneezing, and difficulty breathing. The necropsy found that the affected chicken had obvious pathological changes in the respiratory system and digestive tract, severe congestion and hemorrhage in the tracheal ring, severe congestion and hemorrhage in the lungs, redness, swelling and hemorrhage in the lymph nodes of the small intestine, and enlargement of the liver and kidneys. Acute death occurred in some sick chickens, and the use of various antibiotics did not improve significantly.

[0032] 2. Collection and processing of disease materials

...

Embodiment 2

[0055] Embodiment 2: Preparation of H9 subtype avian influenza X-13 strain virus antigen

[0056] 1. Preparation of seed batches of H9 subtype avian influenza X-13 strain

[0057] Get the purified H9 subtype avian influenza X-13 strain with sterilized normal saline for 10 -4 Doubling dilution, inoculate 9-10 days old SPF chicken embryos, inoculate 0.1mL in the allantoic cavity of each chicken embryo, seal with wax after inoculation, incubate in a 37°C incubator, irradiate embryos once every 8h, remove dead chicken embryos within 24h, Observed to 96h. Harvest the allantoic fluid of chicken embryos in a sterilized container, and measure the hemagglutination value. Any chicken embryos with a titer lower than 7log2 should be discarded. At the same time for sterility test. The harvested virus liquid should be frozen below -30°C before inactivation, which should not exceed 3 months.

[0058] 2. Sterility test

[0059] According to "The Veterinary Pharmacopoeia of the People's R...

Embodiment 3

[0066] Embodiment 3: the immune effect of H9 subtype avian influenza X-13 strain

[0067] 1. Serological potency test

[0068] Fifty 7- to 10-day-old SPF chickens were randomly divided into two groups, 30 in the experimental group and 20 in the control group. The experimental group was subcutaneously injected with the vaccine in the neck, and the dose of immunization preparation was 0.3 mL, among which the dose of antigen was 75 μl. Blood was collected once before inoculation and once a week after inoculation until the fifth week. Blood was randomly collected from 5 mice in each group each time, the serum was separated, and the anti-avian influenza virus H9 subtype HI antibody was detected, and the geometric mean value was taken as the result. The HI detection results are shown in Table 3. The experimental group reached its peak 28 days after immunization, and then maintained at a high level.

[0069] Table 3 Serological potency test

[0070]

[0071] 3. Challenge cross...

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Abstract

The invention discloses a H9 subtype lowly pathogenic avian influenza virus which is classified and named as H9 subtype lowly pathogenic avian influenza virus X-13 strain which is preserved in the China Center for Type Culture Collection; the preservation constitute is called as CCTCC for short; the address of the preservation constitute is Wuhan University, Wuhan, China; the preservation date isJuly 26, 2017; and the preservation number is CCTCC NO:V201733. The H9 subtype lowly pathogenic avian influenza virus disclosed by the invention is good in specificity and immunogenicity, chicken immunized with the strain can endure infection of the H9 subtype lowly pathogenic avian influenza virus. As a vaccine strain for the novel H9 subtype lowly pathogenic avian influenza virus, the strain iscapable of preventing H9 subtype lowly pathogenic avian influenza of livestock, and can be also applied to virus identification and epidemiological investigation.

Description

technical field [0001] The invention belongs to the technical field of biological vaccines, and in particular relates to an H9 subtype low-pathogenic avian influenza virus and its application. Background technique [0002] Avian Influenza (Avian Influenza, AI), referred to as bird flu, is an acute, febrile, highly contagious infectious disease caused by avian influenza virus (Avian Influenza Virus, AIV). Avian influenza viruses belong to the family Orthomyxoviridae, the genus Influenzavirus, and type A avian influenza viruses. Virus particles have a broad spectrum of infection and are pleomorphic and multihost. According to the different pathogenicity and virulence, avian influenza can be divided into three categories, namely highly pathogenic avian influenza, low pathogenic avian influenza and apathogenic avian influenza. Low-pathogenic avian influenza is mainly subtype H9N2, and the clinical symptoms of H9N2 infection are mainly respiratory symptoms, such as: tracheal ed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00G01N33/569A61K39/145A61P31/16C12R1/93
Inventor 李新生黄宗梅崔沛周云飞徐耀辉王洁琼赵玉杰岳旭龙
Owner HENAN AGRICULTURAL UNIVERSITY
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