Avian adenovirus strain, inactivated vaccine and preparation method

A technology of avian adenovirus and inactivated vaccine, which is applied in the field of bioengineering to achieve the effects of high safety, high protection rate against virus and rapid immune response

Active Publication Date: 2019-12-31
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In terms of diagnosis, the current diagnosis is mainly based on clinical symptoms, histopathological changes, PCR amplification, and virus isolation; in terms of disease prevention and control, although there have been research reports on the use of chicken embryos and poultry-derived cells to culture avian adenoviruses, However, the serotypes of most of the reported strains are inconsistent with the current market strains. Therefore, the current market urgently needs to develop a poultry gland with a serotype consistent with the current market strains, high titer, and good immunogenicity. virus vaccine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Preparation of avian adenovirus inactivated vaccine using chicken embryo kidney cells (CEK cells)

[0040] The formula of the cell growth solution is: M199 solution with a volume percentage of 90%, 10% newborn calf serum, and adjust the pH to 7.2;

[0041] The cell maintenance solution formula is: M199 solution with a volume percentage of 98%, 2% newborn calf serum, 2.4mmol / L arginine, and adjust the pH to 7.4;

[0042] a. Preparation of CEK cells: Take 17-day-old SPF chicken embryos, take out the kidneys in a sterile environment, place them in an appropriate amount of pancreatin, and digest them at 37°C. After the digestion is complete, take the supernatant and add an appropriate amount of serum to stop the digestion, centrifuge at 1500 rpm for 10 minutes, discard the supernatant, resuspend the cell pellet with cell growth fluid and dilute to a final concentration of 1×10 cells 6 Cells / ml, cultured at 37°C for 72 hours, the cells grow into a good monolayer;

[0043] ...

Embodiment 2

[0053] Example 2 Preparation of avian adenovirus inactivated vaccine using chicken embryo fibroblast cell line (DF1 cells)

[0054] The growth solution formula for cells is: DMEM / F12 solution with a volume percentage of 92%, 8% newborn calf serum, and adjust the pH to 7.2;

[0055] The cell maintenance solution formula is: DMEM / F12 solution with a volume percentage of 97%, 3% newborn calf serum, 2.2mmol / L arginine, and adjust the pH to 7.3;

[0056] a. Cell culture:

[0057] The DF1 seed cells were removed from the liquid nitrogen and melted in a 37°C water bath, and resuscitated and cultured with cell growth medium (DMEM / F12 medium). After 3 days of culture, the number of cells was 4.0×10 7.0 Cells / ml, and passage 1:3, culture at 37°C for 2 days;

[0058] b. Virus inoculation: Press 10 for poultry adenovirus 3.0 ELD 50 / 0.1ml inoculate DF1 cells, add cell maintenance medium (DMEM / F12 medium), continue culture at 37°C;

[0059] c. Harvesting the venom: After the virus inoculated cells an...

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Abstract

The invention provides a fowl adenovirus strain, an inactivated vaccine and a preparation method. The fowl adenovirus strain is named a fowl adenovirus TJ strain with a preservation number of CCTCC (China Center For Type Culture Collection) NO:V201632; the adenovirus strain is of race C in a fowl adenovirus I subgroup, the serotype is 4 type, viral nucleic acid is double stranded DNA (Deoxyribonucleic Acid), and no envelope is found; the diameter of virus particles is 70 to 90nm, and the virus particles are of icosahedral symmetrical structures; the inactivated vaccine is prepared by performing cell inoculation, venom harvesting, inactivating and emulsifying on a fowl adenovirus; a vaccine strain is a prevalent strain, and is good in immunogenicity and high in multiplication capacity; the prepared vaccine has the advantages of high safety, good stability, quick production of immune response, high virus challenge protecting rate, and the like.

Description

Technical field [0001] The invention relates to a virus strain and an inactivated vaccine, in particular to an avian adenovirus strain, an inactivated vaccine and a preparation method, and belongs to the technical field of biological engineering. Background technique [0002] Chicken Inclusion Body Hepatitis-Hydroperioadium Syndrome (IBH-HPS) is a recently prevalent, acute and highly contagious chicken infectious disease, and its pathogen is Fowl adenovirus (FADV) , In the virus classification, it is Adenoviridae, avian adenovirus of the I subgroup of the avian adenovirus genus. The virus mainly infects broiler chickens, 817 hybrid broiler chickens, and hemp chickens aged 1-3 weeks. It is also more common in broiler breeders and laying hens. Among them, broiler chickens aged 3-7 weeks are more affected. [0003] The disease has spread nationwide in China, mainly in Shandong, Henan, Anhui, Liaoning, Jilin, Hebei, Jiangsu and other places. The disease is mainly transmitted vertical...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/235A61P31/20
CPCA61K39/12A61K2039/5252A61K2039/552C12N7/00C12N2710/10234
Inventor 王帅韩佳丽郁宏伟刘涛何平有朱秀同杨保收梁武刘超
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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