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Method for proliferating and culturing duck tembusu virus by using mosquito-derived C6/36 cell line

A technique for duck tembusu virus and cell culture, which is applied in the directions of microorganism-based methods, viruses, viruses/phages, etc., to achieve the effect of efficient preparation

Inactive Publication Date: 2018-03-23
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the PCR test results showed that the DEF cell culture fluid after inoculation were all positive

Method used

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  • Method for proliferating and culturing duck tembusu virus by using mosquito-derived C6/36 cell line
  • Method for proliferating and culturing duck tembusu virus by using mosquito-derived C6/36 cell line
  • Method for proliferating and culturing duck tembusu virus by using mosquito-derived C6/36 cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0022] Utilize the method for breeding duck Tembusu virus of mosquito source C6 / 36 cell line, concrete steps are as follows:

[0023] First, take out the C6 / 36 cell line (about 1 mL, purchased from Shanghai Hongshun Biotechnology Co., Ltd.) stored in liquid nitrogen, quickly thaw it in a water bath at 37°C within 1 min, and slowly add it to the 9 mL with a pipette. In a 15mL centrifuge tube containing DMEM / F-12 (HAM) cell growth medium (containing 10% fetal bovine serum by volume), centrifuge at 1000r / min for 5min, and discard the supernatant;

[0024] Then add 5 mL of 10vt% fetal calf serum DMEM / F-12 (HAM) cell growth solution to the centrifuge tube, and gently blow and mix with a pipette;

[0025] Move the mixed cell suspension to 25cm 2 Place the cell culture flask at 27°C in 5% CO 2 Cultivate in an insect cell incubator to form a single layer of adherent cells, and end when the cell growth density, that is, the compactness of cells per unit volume or area reaches about 8...

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Abstract

The invention relates to the field of virus culture, and particularly provides a method for proliferating and culturing a duck tembusu virus by using a mosquito-derived C6 / 36 cell line. The mosquito-derived C6 / 36 cell line is used for culturing the tembusu virus; after the method is used, the tembusu virus can be stably proliferated after the tembusu virus is continuously transferred on a C6 / 36 cell for 4-5 generations; meanwhile, obvious cytopathy is formed after inoculating the tembusu virus for 2-3 days.

Description

technical field [0001] The invention relates to the field of virus culture, and specifically provides a method for multiplying and cultivating duck Tembusu virus by using a mosquito-derived C6 / 36 cell line. Background technique [0002] In recent years, my country's duck breeding industry has achieved remarkable development. The number of ducks in stock is about 4 billion, ranking first in the world. However, since the spring of 2010, many duck farms in the southeastern coastal areas of my country (such as Fujian, Shandong, Zhejiang, Shanghai, and Jiangsu, etc.) have broken out a disease characterized by a sharp decline in egg production of laying ducks and breeding ducks, and follicular hemorrhage. Characteristic acute infectious disease. In 2010 alone, about 120 million laying ducks and 15 million meat ducks were infected, causing significant economic losses to my country's poultry farming industry. The pathogen that caused such a serious loss has been confirmed by resear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12R1/93
CPCC12N7/00C12N2770/24151
Inventor 刁有祥唐熠于观留陈浩王琰
Owner SHANDONG AGRICULTURAL UNIVERSITY
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