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Method for preparing duck tembusu virus inactivated vaccine and duck tembusu virus inactivated vaccine

A technology of duck Tembusu virus and inactivated vaccine, applied in the biological field, can solve the problems of large difference between antigen batches, low degree of automation, low virus content, etc., and achieve good safety, high immune efficacy, and high virus content. Effect

Active Publication Date: 2018-11-02
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the disadvantages of high labor intensity, high cost, complex process, low degree of automation, large differences between antigen batches and low virus content in existing cell cultures in the production of duck embryos, a duck embryo stem cell line EB66 is used to produce duck eggs. Method and vaccine for Busu virus disease inactivated vaccine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A method for preparing duck Tembusu virus disease inactivated vaccine, said cell line is duck embryonic stem cell line EB66.

[0037] The above method comprises the steps of:

[0038] 1) Breeding strains: Duck Tambusu virus (Duck Tambusu virus-HB strain, Institute of Animal Husbandry and Veterinary, Beijing Academy of Agriculture and Forestry Sciences, isolated, identified and preserved, gene accession number is JF523187) duck embryo-adapted strain in EB66 cells The cell line (the cell line was purchased by Gansu Jianshun Biology from Valneva Company in France in 2015, and has the exclusive agency right in China) was continuously passaged to the virus-adapted cells, and the chicken embryo limited dilution cloning method (that is, the virus liquid was serially diluted 10 times) , take the virus harvested after inoculating chicken embryos with the highest dilution of venom), take the adapted strain with the highest virus content and continue to culture EB66 cells, and cul...

Embodiment 2

[0044] Duck Tembusu virus disease inactivated vaccine (HB strain) prepared by the method used in Example 1 (appearance milky white homogeneous emulsion; dosage form water-in-oil type. Get a clean straw, absorb a small amount of vaccine, drop it on the clean cold water surface, Except for the first drop, it should not spread; the viscosity should be tested according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and it should meet the requirements; Stability Draw 10mL of the vaccine into a centrifuge tube, centrifuge at 3000r / min for 15 minutes, there should be no stratification, The water precipitated at the bottom of the tube should not exceed 0.5mL; the sterility test should be carried out according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and the growth should be sterile; for the safety test, 5 28-day-old DHOV antibody-negative healthy Peking ducks (or Cherry Valley ducks) were taken. , subcutaneous injection of 1mL in each neck....

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Abstract

The invention discloses a method for preparing a duck tembusu virus inactivated vaccine and the duck tembusu virus inactivated vaccine. A cell line used for virus inoculation is EB66 cell line (a duckembryonic stem cell-derived cell strain), and a bioreactor is adopted for virus culture in a serum-free full suspension manner; and the method comprises the following steps: 1) breeding of virus species; 2) establishment of virus seed batches; 3) preparation of cell venom; 4) inactivation of viruses; and 5) emulsification and other steps to complete the preparation of the vaccine. The method provided by the invention has the characteristics that the prepared cell venom is high in virus content, the production process is stable, intelligent control is achieved, large-scale serum-free suspension culture is realized, the operation is easy, and the cost is low; and the prepared duck tembusu virus inactivated vaccine has the advantages of safety, low side reactions, high immune efficacy, smallbatch-to-batch difference, small number of tests, low cost and the like, and is an ideal vaccine for preventing the occurrence and the prevalence of a duck tembusu virus disease in the waterfowl industry.

Description

technical field [0001] The invention relates to a preparation method of a vaccine, in particular to a method for producing an inactivated vaccine of duck Tembusu virus disease by using a cell line, and belongs to the field of biotechnology. technical background [0002] The number of ducks in our country is about 4 billion per year. Since the spring of 2010, a disease characterized by the decline of duck egg production has broken out successively in Zhejiang, Jiangsu, Shandong, Hebei and Beijing. The ducklings have depression, weight loss, supine, and sideways in the later stage of the disease. If the bacterial secondary infection causes perihepatitis, pericarditis, and air sacculitis, the death rate will increase. In 2010, the disease was named duck hemorrhagic ovariitis (DHO) according to the decline in egg production and follicular hemorrhage. Later, etiological research confirmed that the disease was caused by DuckTembusu Virus (DTMUV) of the flavivirus family. Caused,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/14C12N7/06C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61P31/14C12N7/00C12N2770/24134C12N2770/24163
Inventor 刘月焕罗顺林健张业炘杨志远王嘉琪王小蕾杨云段会娟杨贵君刘立新耿风廷程慧敏陈佩兰赵际成赵志欣潘洁钱建宁王盟
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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