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A method for screening virus-sensitive cell clones using indirect immunofluorescence detection technology

A technology for immunofluorescence detection and virus-sensitive cells, which is applied in the biological field, can solve the problems of long cycle, high cost, and complicated operation, and achieve the effect of strong repeatability, short detection time, and simple operation

Active Publication Date: 2015-10-07
山东滨州沃华生物工程有限公司
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Problems solved by technology

However, for non-pathogenic viruses, the determination of the toxicity can only be carried out by using animals or other methods, such as the determination of the attenuated virus titer of classical swine fever rabbit
This method is time-consuming, labor-intensive, complicated to operate, long in cycle, high in cost and low in screening efficiency

Method used

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  • A method for screening virus-sensitive cell clones using indirect immunofluorescence detection technology
  • A method for screening virus-sensitive cell clones using indirect immunofluorescence detection technology
  • A method for screening virus-sensitive cell clones using indirect immunofluorescence detection technology

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Embodiment Construction

[0043] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention, but the present invention is not limited thereby.

[0044] (1) Identification of cell line purity:

[0045] Recovery culture of ST cells: Culture ST cells recovered from liquid nitrogen in a square bottle. The culture conditions include: pH 7.2, temperature 37°C, medium MEM containing 8% serum for swine fever; culture time 72h, cell single Layer length to 100%;

[0046] Subculture of ST cells: Digest and disperse the monolayer of ST cells with EDTA-trypsin to obtain a cell suspension, which is divided into square flasks according to the culture area ratio of 1:3;

[0047] Inoculation of cells: attenuated hog fever rabbit virus (preserved by the China Veterinary Drug Administration, preservation number AV1412) stored at -80°C (poison price 10 3.0 TCID 50 / 0.1mL) in wa...

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Abstract

The invention relates to the field of biotechnology, and particularly relates to a method for performing screening in virus-sensitive cell clonal strains by applying an indirect immunofluorescence assay technology. The method comprises the following steps of: identifying the purity of a cell line; obtaining monoclonal cell strains in the cell line; and identifying the sensitivity of the monoclonal cell strains to virus. The method disclosed by the invention has the beneficial effects of being short in detection time, capable of identifying the sensitivity of single-cell cloning to virus only by 3 days, relative simple to operate, easy, capable of being used for screening lots of non-cytopathic virus-sensitive cell strains, stable in result, strong in repeatability, and consistent with the effect of a virulence determination method.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a method for screening virus-sensitive cell clones using indirect immunofluorescence detection technology. Background technique [0002] Traditionally, the screening of sensitive cell lines is achieved by measuring the virus titer. However, for non-pathogenic viruses, the determination of the toxicity can only be carried out by using animals or other methods, such as the determination of the attenuated toxicity of classical swine fever rabbit. This method is time-consuming, labor-intensive, complicated to operate, long in cycle, high in cost and low in screening efficiency. Contents of the invention [0003] In view of the above deficiencies, the present invention provides a simple and quick screening method for sensitive cell lines, specifically, the identification of the purity of cell lines and the sensitivity identification of different monoclonal cell lines by...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071G01N33/53
Inventor 陈申秒董新荣王常伟徐海军王楠
Owner 山东滨州沃华生物工程有限公司
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