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Method for producing porcine reproductive and respiratory syndrome viruses (PRRSV) in large scale

A technology of porcine PRRS virus and PRRS virus, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem of limited product yield, improvement of quality, reduction of production cost, and short cell maintenance time , Intensive manual operation and other issues, to achieve the effect of saving cost and manpower, improving immunity, and high efficiency of virus infection

Inactive Publication Date: 2010-09-15
PU LIKE BIO ENG
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Problems solved by technology

[0004] The existing large-scale production of PRRS virus mostly adopts the spinner bottle process, but the manual operation is intensive in the production process, the virus strain is limited, the cell maintenance time is short, the density and activity are low, etc., which limit the improvement of the output and quality of the product and production cost reduction

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  • Method for producing porcine reproductive and respiratory syndrome viruses (PRRSV) in large scale

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Embodiment 1

[0031] 1. Viruses and cell lines

[0032] The virus strain used to prepare the PRRS virus antigen is the NVDC-JXA1 strain, and the pathogenicity test shows that the virus of the strain is not pathogenic. The monkey kidney cell (Marc-145), a cell line with good sensitivity to the virus strain, was used as a cell line for seedling production, so as to infect and multiply the PRRS virus in large quantities.

[0033] 2. Preparation method

[0034] (1) Inoculate the monkey kidney cells (Marc-145), the cell line used for the preparation of the antigen, into the MEM liquid medium (including 10% bovine serum, 0.01mol / L NaHCO 3 , 0.1mg / ml kanamycin sulfate (kanamycinsulfate), 100,000IU penicillin G sodium salt (Penicillin G Sodium), pH value is 7.2) and in the carrier tank of BioNoc II polyester fiber; Add 5.5g of microcarriers, the initial inoculum size of cells is 8.0×10 9 cells / g microcarriers.

[0035] (2) At 37°C, 5% CO 2 Under the culture environment, the cells are attached ...

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Abstract

The invention discloses a method for producing porcine reproductive and respiratory syndrome viruses (PRRSV) in a large scale. The method comprises the following steps of: producing PRRSV by using a cell microcarrier suspension culture system by utilizing a bioreactor; inoculating host cells used for preparing the viruses to a carrier pot containing a culture solution and microcarriers and uniformly mixing the cells and the microcarriers to ensure that the cells are attached on the microcarriers; under a proper culture environment, providing sufficient nutrients and an appropriate gas environment for the cells to ensure that the cells grow on the microcarriers to reach a concentration 5-40 times of an inoculating concentration; changing to use a cell maintenance culture solution and preparing the PRRSV into a viral suspension to ensure that the viral suspension is adsorbed on the cells; culturing the viruses under an appropriate environment; obtaining a viral solution after continuously culturing for 2-3 days; after the viral solution is inspected to be qualified, freeze thawing the obtained viral solution twice at -20 DEG C; and inactivating and purifying to obtain a PRRSV solution. The method has large production scale, high single-batch output and relatively lower production cost.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, relates to animal virus culture technology, and more specifically relates to a method for producing pig PRRS virus. Background technique [0002] Pig blue-ear disease, also known as porcine reproductive and respiratory syndrome, is a viral infectious disease that endangers the pig industry. In the summer and autumn of 2006, pig "high fever" epidemics occurred in some provinces in southern my country. The Ministry of Agriculture actively organized scientific research personnel to jointly tackle the problem, and identified the pig PRRS variant virus as the main pathogen, which was named "highly pathogenic pig PRRS". Until today, the epidemic situation is still severe. Like other viral diseases, there is currently no effective therapeutic drug for the treatment of PRRS. Immunization is the most effective way to prevent and control the disease. Currently commercialized PRRS Va...

Claims

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Application Information

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IPC IPC(8): C12N7/04C12N7/02C12R1/93
Inventor 张许科孙进忠乔荣岑陶家权骆爱芳张海洋习向锋李三
Owner PU LIKE BIO ENG
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