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Production of avian reovirus and vaccine by using mammalian cell line

A technology of poultry reovirus and mammals, applied in the field of veterinary vaccines, can solve the problems of complicated process, high cost and high vaccine price

Active Publication Date: 2013-07-03
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main means of controlling avian reovirus in my country is to use attenuated live vaccines or whole virus inactivated vaccines to control chicken reovirus infection through immunization, while the avian reovirus vaccines currently used in my country are all foreign countries Imported vaccines use chicken embryos or chicken embryo fibroblasts to culture and produce viruses, which makes the vaccines expensive and the process is complicated, resulting in higher costs for farmers to prevent and control ARV

Method used

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  • Production of avian reovirus and vaccine by using mammalian cell line
  • Production of avian reovirus and vaccine by using mammalian cell line

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1: Using Vero cells to prepare avian reovirus and avian reovirus inactivated vaccine

[0023] (1) Selection of cells for seedling production: African green monkey kidney (Vero) cells were selected and purchased from Shanghai Institute of Biochemistry.

[0024] (2) Passage and culture of cells for seedling production: Vero cells were digested and passaged with EDTA-trypsin cell dispersion solution, and continued to culture at 37°C with cell growth medium. When a good monolayer was formed, they were used for continued passage or virus inoculation.

[0025] (3) Propagation of cytotoxic species:

[0026] Preparation of chicken embryo kidney cells (CK cells): Take 10 17-20-day-old SPF chicken embryos, cut open the abdominal cavity, remove the internal organs, carefully separate the capsule of the kidney with tweezers, then take out the kidney, wash it 3 times with Hank's solution, Then cut it to 1mm 3 Add an appropriate amount of 0.25% trypsin to the cube, and d...

Embodiment 2

[0036]Embodiment 2: Comparison of test results between cell-derived ARV inactivated vaccine and chicken embryo-derived ARV chicken embryo-derived inactivated vaccine

[0037] 1 material

[0038] 1.1 Vaccines Three batches of ARV inactivated vaccines (cell line source) were prepared according to Example 1, batch numbers: Trial Vero20110301, Trial Vero20110302, Vero20110303. There are 3 batches of chicken embryo sources, batch numbers: 201101, 201102, 201103, and the preparation method is shown in 2.1 of this example.

[0039] 1.2 SPF eggs and SPF chickens were purchased from Beijing Meria Weitong Biotechnology Co., Ltd.

[0040] Preparation method and results of 2ARV chicken embryo-derived inactivated vaccine

[0041] 2.1 Preparation of chicken embryo-derived ARV inactivated vaccine:

[0042] a. Virus propagation: AV2311 strain virus seeds are propagated and passed on chicken embryos to make the virus titer detection reach 10 10.5 EID 50 / 0.2ml; b. vaccine inactivation: fo...

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Abstract

The invention provides a production method for avian reovirus by using a mammalian cell line. The production method comprises the following steps: using a mammalian cell line like a Vero cell line or an MDCK cell line which has formed a cell monolayer with good growth, inoculating avian reovirus which undergoes subculture by using avian kidney cells and allowing avian reovirus to be adsorbed on the cell line; and culturing avian reovirus in a cell maintenance medium instead so as to realize propagation of avian reovirus and harvesting the avian reovirus. An inactivator is added into the avian reovirus produced by using the method, then a vaccine adjuvant acceptable in veterinary pharmacy is added after inactivation, and finally, emulsification is carried out so as to obtain an avian reovirus vaccine.

Description

technical field [0001] The invention relates to a method and product of avian reovirus and vaccine produced by mammalian cell lines, belonging to the field of veterinary vaccines. Background technique [0002] Avian Reovirus (ARV) exists widely in the world and is widely prevalent in chickens and turkeys. The arthritis, tenosynovitis and immunosuppression it causes have brought significant economic losses to the chicken industry. At present, the main means of controlling avian reovirus in my country is to use attenuated live vaccines or whole virus inactivated vaccines to control chicken reovirus infection through immunization, while the avian reovirus vaccines currently used in my country are all foreign countries Imported vaccines use chicken embryos or chicken embryo fibroblasts to culture and produce viruses, which makes the vaccines expensive and complicated, resulting in increased costs for farmers to prevent and control ARV. Contents of the invention [0003] In vie...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/02A61K39/15A61P31/14
CPCA61K2039/5252A61K2039/552C12N2720/12234C12N2720/12251
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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