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Method for eliminating mycoplasma contamination of PCV2 (Porcine Circovirus Virus 2)

A mycoplasma and virus technology, applied in the direction of virus/bacteriophage, biochemical equipment and methods, microorganisms, etc., can solve the problem that the complete removal of mycoplasma PCV2 virus titer cannot be guaranteed, and achieve the removal of mycoplasma in a short time, small concentration, and virus content guarantee Effect

Active Publication Date: 2014-10-01
HANGZHOU JIANLIANG VETERINARY BIOLOGICAL PREPARATIONS CO LTD
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Problems solved by technology

Therefore, the single use of any of the above methods cannot guarantee the complete removal of mycoplasma while ensuring that the titer of the PCV2 virus will not decrease

Method used

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  • Method for eliminating mycoplasma contamination of PCV2 (Porcine Circovirus Virus 2)
  • Method for eliminating mycoplasma contamination of PCV2 (Porcine Circovirus Virus 2)

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Embodiment Construction

[0038] Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail:

[0039] A kind of method that the invention provides to remove mycoplasma pollution in PCV2 virus, concrete steps comprise:

[0040] (1) 0.1μm bacteria filter filters PCV2 virus and reproduces it for four generations;

[0041] (2) PCV2 virus with a final concentration of 2.5μ g / ml Plasmocin treatment for two generations;

[0042] (3) Two generations of conventional breeding of PCV2.

[0043] Described step (1) specifically comprises the following steps:

[0044] A. Cultivate PK-15 cells with MEM medium (GIBCO, 574131) containing 5% fetal bovine serum (PAA, A10209-1611), and the cell density is 2×10 5 PCV2 virus is filtered with a 0.1 μm bacteria filter, and the PCV2 virus is inoculated in the MEM medium, so that the PCV2 virus inoculation amount is 1% of the volume of the MEM medium, and then the MEM medium containing the PCV2 virus is inocu...

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Abstract

The invention relates to the technical field of biology, and aims to provide a method for eliminating mycoplasma contamination of PCV2 (Porcine Circovirus Virus 2). The method for eliminating the mycoplasma contamination of the PCV2 comprises the following steps: filtering the PCV2 through a 0.1-micrometer bacteria filter, and breeding for 4 generations; treating two generations of the PCV2 by using Plasmocin with the final concentration of 2.5 micrograms / milliliter, and conventionally breeding the PCV2 for two generations. The method provided by the invention saves the experiment cost and more importantly, has the advantages of few antibiotic usage times, low concentration and short mycoplasma removing time by adopting a method of firstly filtering and then using antibiotics and can complete the integral treatment process for only 8 generations and 16-20 days without lowering the titer of the PCV2 viruses, provide the mycoplasma-free and high-titer PCV2 viruses for vaccine production and provide the guarantee for the increase of the virus content of a vaccine semi-finished product within unit volume.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for removing mycoplasma pollution in PCV2 virus. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) belongs to the family Circoviridae, the genus Circovirus, has no envelope, and is composed of a single strand of covalent closed circular DNA. The diameter of the virus particle is about 17nm, and it is icosahedral. It is the smallest vertebrate virus ever discovered. The virus can be divided into two serotypes, PCV1 and PCV2. Among them, PCV1 has no pathogenicity to pigs, and is widely present in pigs and pig-derived cell lines. PCV2 can cause a variety of infectious diseases in pigs, such as multisystemic wasting syndrome (PMWS) in weaned piglets, porcine dermatitis and nephropathy syndrome. sign (PDNS) and so on. PCV2 can not only cause slow growth of piglets, but also infringe on the immune system of pigs, leading to immunosuppression in pig herd...

Claims

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Application Information

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IPC IPC(8): C12N7/02C12N7/00
Inventor 李少丽
Owner HANGZHOU JIANLIANG VETERINARY BIOLOGICAL PREPARATIONS CO LTD
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