Group I avian adenovirus type 8 strain and application thereof

A technology of poultry adenovirus and strains, applied in the application field of bivalent inactivated vaccines, can solve the problems of wrong type of cross-protection vaccine and poor protection effect, and achieve long duration of immunity, stable immune effect and good safety Effect

Active Publication Date: 2019-01-15
乾元浩生物股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, some studies have shown that the cross-protection between different serotypes of avian adenoviruses is weak, resulting in wrong types of vaccines, and the protection effect is not good. Therefore, the key to vaccine development lies in the development of corresponding serotypes of avian adenoviruses that are prevalent in the area where the vaccine is used. Products, it is very important to do a good job in the prevention and control of the type
So far, there is no commercially available bivalent inactivated vaccine of avian adenovirus serotype 4 and serotype 8

Method used

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  • Group I avian adenovirus type 8 strain and application thereof
  • Group I avian adenovirus type 8 strain and application thereof
  • Group I avian adenovirus type 8 strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Isolation and Screening of Virus Strains

[0041] 1. Isolation and identification of avian adenovirus strains

[0042] Since 2014, the inventor has collected multiple suspected poultry adenovirus-infected tissue samples in Liaoning, Shandong, Henan, Jiangsu and other places. The main apparent necropsy lesions of the sick chickens: it can be seen that about 5-10mL of adenovirus has accumulated in the pericardial cavity. Light yellow clear water or jelly-like liquid; the liver is lighter in color, with local necrotic lesions, and the kidneys are pale, swollen, and bleeding.

[0043]Collect the liver and spleen tissues of diseased chickens with typical symptoms and pathological changes, cut them into pieces and grind them, add sterilized PBS at a weight ratio of 1:3 after weighing, centrifuge at 4°C, 6000r / min for 10min, and centrifuge at 4°C, 8000r / min. Centrifuge for 10 min, transfer the supernatant to another sterile 1.5 mL centrifuge tube, add double antibod...

Embodiment 2

[0051] Example 2 ZMXZAV-4 strain, ZMYTAV-8 strain identification and properties

[0052] 1. Sterility test

[0053] Tested according to the appendix of the current "Chinese Veterinary Pharmacopoeia", ZMXZAV-4 strain and ZMYTAV-8 strain F1 generation seed virus samples grew aseptically after inoculation respectively.

[0054] 2. Determination of virus content

[0055] After ZMXZAV-4 strain, ZMYTAV-8 strain F1, F5, F10, F15 generation virus solution was diluted 10 times, take 10 -5 、10 -6 、10 -7 、10 -8 、10 -9 、10 -10 Six dilutions were inoculated into well-growing 96-well culture plates of LMH cells, and placed at 37°C and 5% CO 2 After 2 hours of incubation, maintenance solution was added and culture continued for 168 hours. After 168 hours, the cell lesions were observed hole by hole, and the typical lesions of the inoculated cells were judged as infection, and the virus content was calculated according to the Reed-Muench method. The results show that the virus conten...

Embodiment 3

[0072] Embodiment 3 ZMXZAV-4 strain, ZMYTAV-8 strain virus seed preparation

[0073] 1. Optimal inoculum volume

[0074] ZMXZAV-4 strain and ZMYTAV-8 strain seed virus were inoculated on well-growing monolayer LMH cells according to 1:100, 1:1000, and 1:10000 respectively. Each sample was repeated 3 times, and placed at 37 ° C, 5% CO 2 Cultivate and observe the cell changes. When more than 80% of the cells are damaged, harvest the virus solution. After repeated freezing and thawing twice, use the real-time PCR method to measure the virus particles in the culture, and select the one with the highest concentration to determine the optimal inoculation dose.

[0075] 2. The best time to harvest

[0076]According to the results of the optimal inoculation dose test, select the optimal inoculation dose, inoculate on the well-growing monolayer LMH cells, freeze and thaw after 24, 48, 72, 96, 120, 144, 168, and 192 hours after inoculation, respectively Three times, the cell culture w...

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Abstract

The invention relates to the technical field of veterinary biological products, in particular to a group I avian adenovirus type 8 strain and application thereof. The group I avian adenovirus type 8 strain ZMYTAV-8 strain, with an accession number of CGMCC No. 14297 has the characteristics of high toxin production and good immunogenicity. The strain can effectively prevent chicken inclusion body hepatitis caused by serum avian adenovirus type 8. Therefore, the invention also provides application of the avian adenovirus type 8 strain ZMYTAV-8 in the preparation of drugs for the prevention of inclusion body hepatitis. Further, the present invention provides a bivalent inactivated vaccine of avian adenovirus serotype 4, serotype 8 for the prevention of pericardial effusion-hepatitis syndromeand inclusion body hepatitis. The vaccine has good immunogenicity and can prevent avian diseases caused by adenovirus, such as inclusion body hepatitis and pericardial effusion-hepatitis syndrome, which are popular in recent years. The vaccine has 100% protection rate against local isolates, and has high safety, rapid antibody production and stable efficacy.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a group I type 8 avian adenovirus strain and its application in a bivalent inactivated vaccine of virus serotype 4 and serotype 8. Background technique [0002] Avian adenovirus can be divided into 5 species and 12 serotypes: A, B, C, D, E, which can cause various diseases in chickens, ducks, geese and other poultry or wild birds. The main clinical manifestations include pericardial effusion and inclusion bodies Hepatitis, aplastic anemia, hemorrhage, mild respiratory disease and decreased egg production, etc. [0003] Avian adenovirus has strong resistance to the external environment, and its complex transmission mode makes it spread and become popular in recent years, causing great impact and loss to the domestic poultry breeding industry. Especially since 2014, the scope of its influence has continued to expand, and local epidemic cases have been report...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/295A61K39/235A61P1/16A61P31/20C12R1/93
CPCA61K39/12C12N7/00A61K2039/70A61K2039/5252A61K2039/552C12N2710/10221C12N2710/10234
Inventor 张贺楠张钊伟于雷邵葳周建民倪娇赵成全
Owner 乾元浩生物股份有限公司
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