Culture method of porcine pseudorabies virus

The technology of a porcine pseudorabies virus and a culture method is applied in the field of culturing porcine pseudorabies virus by cell culture technology, which can solve the problems such as not easy to grasp the virus, achieve good social benefits and application prospects, complete control effect, and high yield effect

Inactive Publication Date: 2014-06-18
QINGDAO ZHONGREN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In order to overcome the above-mentioned defects existing in the prior art field, the object of the present invention is to provide a method for cultivating porcine pseudorabies virus, which solves the problem that the prior art adopts the method of cell proliferation virus to prepare antigen, and it is not easy to grasp the law of virus and select the best cell. Problems with viral culture

Method used

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Embodiment

[0017] The culture method of the porcine pseudorabies virus of the present embodiment may further comprise the steps:

[0018] (1) Cell subculture, digest BHK-21 cells with trypsin-EDTA solution, after the cells are loose, add DMEM cell culture medium, blow the digested cells evenly with a pipette, pour out 4 / 5 of the cell suspension solution, and then add an equal amount of DMEM cell culture medium, that is, dilute the cell density to 5 times;

[0019] (2) Virus proliferation, inoculate BHK-21 with 2% PRV seed in a monolayer, and set up uninfected normal control cells at the same time, incubate at 37°C for 1 hour, add cell maintenance solution, and place at 37°C, 5% CO 2 Cultivate in an incubator, observe the time and characteristics of CPE appearance, collect the poison when 80% CPE appears in the cells, freeze and thaw 3 times, centrifuge at 1500 r / min for 15 minutes, divide the supernatant into the vials for later use, and pass this method to the third generation , store ...

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Abstract

The invention discloses a culture method of porcine pseudorabies virus; hamster kidney cells (BHK-21) are used as a cell source for culturing the porcine pseudorabies virus, and while cell passage, the virus is inoculated, so that multiplication culture of the virus is realized. The method has the characteristics of simple technology, high increment, high yield and low cost, and a vaccine prepared by use of the porcine pseudorabies virus cultured by the method has a complete preventive effect on the porcine pseudorabies virus, and has good social benefits and application prospects.

Description

[0001] technical field [0002] The invention belongs to the technical field of porcine pseudorabies virus vaccine production, and in particular relates to a method for cultivating porcine pseudorabies virus with cell culture technology. [0003] Background technique [0004] Porcine pseudorabies (PR) is an acute, febrile infectious disease caused by pseudorabies virus (PRV), characterized by fever and encephalomyelitis. The disease can cause pigs, cattle, sheep, dogs, cats, rabbits, mice, minks, foxes and other domestic and wild animals to become ill, and has a high fatality rate. Pigs are the only survivable host for the virus to proliferate and infect, and they are the natural reservoir of the virus. Therefore, cutting off the transmission of pseudorabies virus from pigs is an effective way to control pseudorabies. Adult pigs generally show recessive infection, causing growth stagnation, slow weight gain, etc.; pregnant sows can cause syndromes such as abortion, stillb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12R1/93
Inventor 张述智夏伟朱绍辉张浩王晓丽徐权汗李之详许团辉
Owner QINGDAO ZHONGREN PHARMA
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