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Primer for detecting Vesivirus 2117 viral pollution in CHO cell, kit and method

A kit and virus technology, applied in the field of biomedicine, can solve the problems of dye redistribution, strong amplification signal, etc., and achieve the effect of ensuring reliability

Pending Publication Date: 2019-03-15
河南普诺易生物制品研究院有限公司 +1
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  • Description
  • Claims
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Problems solved by technology

The kit has a strong amplification signal and can solve the problem of dye redistribution

Method used

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  • Primer for detecting Vesivirus 2117 viral pollution in CHO cell, kit and method
  • Primer for detecting Vesivirus 2117 viral pollution in CHO cell, kit and method
  • Primer for detecting Vesivirus 2117 viral pollution in CHO cell, kit and method

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preparation example Construction

[0058] The preparation steps of the positive quality control product are as follows: the specific DNA sequence containing the Vesivirus 2117 virus is cloned into the pUC57 plasmid vector by recombinant DNA genetic engineering method, then transformed into Escherichia coli, and the recombinant plasmid DNA is obtained through steps such as plasmid amplification and extraction, That is, the positive quality control product, the pUC57 recombinant plasmid containing Vesivirus 2117 virus constitutes a schematic diagram as shown in figure 1 shown.

[0059] The embodiment of the real-time fluorescent quantitative PCR method that detects Vesivirus 2117 virus contamination in CHO cells

[0060] The present embodiment detects the real-time fluorescent quantitative PCR method of Vesivirus 2117 virus contamination in CHO cells, comprising the following steps:

[0061] 1) Take the positive quality control as a standard, dilute it into a solution with a concentration of 1ng / μL, and then dil...

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Abstract

The invention relates to a primer for detecting Vesivirus 2117 viral pollution in a CHO cell, a kit and a method and belongs to the technical field of biomedicines. A real-time fluorogenic quantitative PCR primer sequence for detecting Vesivirus 2117 virus is as follows: a forward primer: 5'-ACGTTGACGATAGGGGGAGA-3' and a backward primer: 5'5-CTTTGGCAAAGACCCCGTTG-3'. The kit also comprises a positive quality control and reference housekeeping gene GAPDH primer sequences which are as follows: a forward primer: 5'-CCCATGGCAAGTTCAAAGGCA-3' and a backward primer: 5'-TGGTGAAGACGCCAGTAGATT-3'. The kit has the characteristics of being high in sensitivity, high in specificity and high in efficiency, and is simple to operate and time-saving.

Description

technical field [0001] The invention relates to a primer, a kit and a method for detecting Vesivirus 2117 virus contamination in CHO cells, belonging to the technical field of biomedicine. Background technique [0002] Vesivirus 2117 belongs to the family Caliciviridae, which is one of the contaminants in cell culture operations in the biopharmaceutical industry. Caliciviridae are a ubiquitous family of small round viruses found in marine and some terrestrial mammals. Virions are non-enveloped, 27-40 nm in diameter, and have icosahedral symmetry. Some virions have a characteristic 32-cup appearance, whereas enteric caliciform virions lack characteristic surface decoration and have an indistinct appearance. The RNA genome of Vesiviruses contains three major open reading frames (open reading frames, ORF-1, -2, -3). ORF-1 encodes a proteolytic mature nonstructural protein. ORF-2 encodes the major capsid protein VP1, and ORF-3 encodes the small structural protein VP2. At pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 林重超王建华王天云郭潇杨献军王燕芳
Owner 河南普诺易生物制品研究院有限公司
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