Method for producing marek disease live vaccine of chicken by using cell line

A technology of chicken Marek's disease and cell lines, which is applied in the field of production of chicken Marek's disease live vaccines with cell lines, can solve problems affecting vaccine production and quality, contamination of cells with exogenous viruses, and low vaccine potency, and achieve good economy Benefits and application prospects, high virus content, good immune efficacy

Active Publication Date: 2014-08-13
南京创启生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although my country has stipulated that chicken embryos and cells for preparing poultry vaccines must come from specific pathogen-free (SPF) chicken flocks, in reality, due to the lag of detection and the complexity of the poultry breeding environment in my

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  • Method for producing marek disease live vaccine of chicken by using cell line
  • Method for producing marek disease live vaccine of chicken by using cell line
  • Method for producing marek disease live vaccine of chicken by using cell line

Examples

Experimental program
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Example Embodiment

[0029] Example 1

[0030] Preparation of virus seed: Take out the stored CVI988 / Rispens virus seed from liquid nitrogen, melt it in warm water at 37℃ (within 1 minute), immediately dilute it in DF-1 cell growth medium at 1:5, and centrifuge at 1000r / min. After 10 minutes, discard the supernatant, dilute with cell growth solution to a 100ml square bottle containing 200,000 PFU / ml, inoculate DF-1 monolayer, inoculate 0.5ml per bottle, absorb at 36.5-37.5°C for 1 hour, add maintenance solution Put 5% CO2 at 36.5-37.5°C for 2-4 days. When 70% of the monolayer has cytopathic changes, a large number of confluent cells and refraction round cells are produced, 0.25% pancreatin-0.02% EDTA (1:4) The cell dispersion is digested and dispersed the cells, the harvested cell suspension is centrifuged at 1000 r / min for 10 minutes, the supernatant is discarded, and the precipitated cells are suspended in an appropriate amount of maintenance solution to inoculate the DF-1 monolayer to continue...

Example Embodiment

[0041] Example 2

[0042] Propagation of the virus seed: use the basal seed of Marek’s disease III turkey herpes virus Fc-126 with seed virus dilution (2 times 199 of 5% newborn calf serum and an equal amount of mixed liquid of 2 times hydrolyzed milk protein solution, containing 100 Unit / ml double antibody, pH adjusted to 7.2) Dilute to 400,000 PFU / ml, inoculate 0.5ml / 100ml square flask on well-growing DF-1 cells, adsorb at 36.5-37.5°C for 1 hour, then add appropriate amount of maintenance solution , Continue to culture, when 70% or more of the monolayer cells have typical Marek’s disease cell lesions, digest and disperse the cells with EDTA-trypsin cell dispersion, and the harvested cell suspension is used as a seedling virus seed;

[0043] Subculture and culture of seedling-making cells: DF-1 cell line is digested and subcultured with EDTA-trypsin cell dispersion, and cultured with cell growth solution at 36.5-37.5°C to form a good monolayer, used for continued passage or virus ...

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Abstract

The invention discloses a method for producing a marek disease live vaccine of chicken by using a cell line. The method comprises the following steps: A, selecting the cell line as a vaccine-preparing cell; B, passaging and cultivating the vaccine-preparing cell; C, breeding cytotoxic species; D, breeding vaccine-preparing venom; E, preparing vaccine, carrying out split charging, freeze-drying or cryopreserving in liquid nitrogen. By adopting the method, the defects of the prior art can be solved, the method has the advantages of being stable in production process, easy to operate, high in virus content, small in difference between batches, and easy in quality control, and the yield and quality of the vaccine can be significantly improved. The marek disease live vaccine of chicken produced by the method is good in safety, and high in immunity, and has complete immune protection action on attack of a virulent strain or a very virulent strain of chicken marek's disease.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for producing chicken Marek's disease live vaccine with cell lines. Background technique [0002] The cells used in the production of chicken Marek's disease live vaccines in my country are primary chicken embryo fibroblasts. Although my country has stipulated that chicken embryos and cells for preparing poultry vaccines must come from specific pathogen-free (SPF) chicken flocks, in reality, due to the lag of detection and the complexity of the poultry breeding environment in my country, chicken embryos for poultry vaccine production Cells are easy to cause cell-exogenous virus contamination, and the vaccine potency is not high, and the batch-to-batch variation is large, which seriously affects the yield and quality of the vaccine. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a metho...

Claims

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Application Information

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IPC IPC(8): A61K39/245A61K39/255A61P31/22A61P35/00
Inventor 陆秀云
Owner 南京创启生物科技有限公司
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