Method of screening MDBK (Madin-Darby Bovine Kidney) cell lines sensitive to bovine viral diarrhea/mucosal viruses

A bovine viral diarrhea and cell line technology, which is applied in the field of screening MDBK cell lines sensitive to bovine viral diarrhea/mucosal virus, and achieves the effects of short time, stable results, and simple and easy operation.

Active Publication Date: 2016-07-06
JINYUBAOLING BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the late start in China, insufficient attention has been paid to bovine viral diarrhea/mucosal disease, and there is no commercialized vaccine, which has led to the serious epide...

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  • Method of screening MDBK (Madin-Darby Bovine Kidney) cell lines sensitive to bovine viral diarrhea/mucosal viruses

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Embodiment 1

[0040] Embodiment 1, screening the MDBK cell line sensitive to bovine viral diarrhea / mucosal virus

[0041] Screen the method for the MDBK cell line sensitive to bovine viral diarrhea / mucosal virus with the present invention, can carry out following operations:

[0042] 1) Recovery of MDBK cells

[0043] Take out MDBK cells (commercially available) without exogenous virus contamination from liquid nitrogen, immerse them in warm water at 37°C, and shake them from time to time to melt them as soon as possible, take out the cryopreservation tube, suck out the cell suspension with a straw, and add to 15 mL Add 10 mL of MEM medium (purchased from Gibco) to the centrifuge tube, mix well and centrifuge at 1000 rpm for 5 min, discard the supernatant, and obtain MDBK cells without exogenous virus contamination.

[0044] 2) Culture MDBK cells

[0045] MDBK cells without exogenous virus contamination were resuspended in MEM medium containing 10% (volume / volume, V / V, unit mL / 100mL) feta...

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Abstract

The invention discloses a method of screening MDBK (Madin-Darby Bovine Kidney) cell lines sensitive to bovine viral diarrhea/mucosal viruses. The method includes: performing mono-cloning on MDBK cells; inoculating bovine viral diarrhea/mucosal viruses of two biotypes (cytopathic and noncytopathic); adopting indirect immunofluorescence technology to measure viral titer, and determining the cell lines with the viral titer higher than 106TCID50/ml as the MDBK cell lines sensitive to the bovine viral diarrhea/mucosal viruses. The MDBK cell lines screened by the method are sensitive to the bovine viral diarrhea/mucosal viruses of the two biotypes, and the method can be directly applied in research and production of bovine viral diarrhea/mucosal disease vaccines.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a method for screening MDBK cell lines sensitive to bovine viral diarrhea / mucosal virus. Background technique [0002] Bovine viral diarrhea / mucosal disease (bovineviraldiarrhea, BVD) is an infectious disease of cattle caused by bovineviraldiarrheavirus (BVDV). The causative agent, bovine viral diarrhea / mucosal virus, is a member of the Pestivirus genus of the Flaviviridae family and is a single-stranded RNA, enveloped virus. Bovine viral diarrhea / mucosal virus can be divided into two biotypes according to whether it can cause cytopathic effects on cultured cells: one is non-cytopathic because the virus replicates in cells without causing cytopathic effects; the other is non-cytopathic The virus replicates in cells and causes cells to form vacuoles, undergo nuclear pyknosis, dissolve and die, and is called cytopathic type. [0003] The clinical manifestations of cows inf...

Claims

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Application Information

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IPC IPC(8): C12N5/071G01N33/569
CPCG01N33/56983C12N5/0686G01N2333/183C12N2509/00C12N2500/84
Inventor 商晓桂孔彩萍韩志玲郝鹏羡东堡田志辉王云凌范秀丽刘国英魏学峰
Owner JINYUBAOLING BIO PHARMA CO LTD
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