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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
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  • Claims
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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 epidemic of the disease nationwide. It is urgent to carry out relevant vaccine research to prevent, control and eventually eradicate it in my country. Safe and effective vaccine guarantees for this disease

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

Examples

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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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