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A method for rapid detection of residual bhk-21 cells cultured in foot-and-mouth disease vaccine

A BHK-21, foot-and-mouth disease vaccine technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve accurate results, ensure specificity and reliability, and good repeatability.

Active Publication Date: 2019-02-01
云南省动物疫病预防控制中心
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  • A method for rapid detection of residual bhk-21 cells cultured in foot-and-mouth disease vaccine
  • A method for rapid detection of residual bhk-21 cells cultured in foot-and-mouth disease vaccine
  • A method for rapid detection of residual bhk-21 cells cultured in foot-and-mouth disease vaccine

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Embodiment Construction

[0050] The following is attached Figure 1-3 , the substantive content of the present invention is further illustrated by the embodiments of the present invention, but the present invention is not limited thereto.

[0051] The techniques used in the following examples include nucleic acid extraction and PCR amplification. Molecular biology techniques such as enzyme digestion and identification are all conventional techniques known to those skilled in the art; the instruments, equipment, reagents, cell lines, etc. used are all research and technical personnel in this field, unless otherwise specified in this specification. .

[0052] One, the preparation method of Cytb plasmid standard

[0053] 1. Construction of Cytb plasmid standard:

[0054] The conserved sequence of the Cytb gene of BHK-21 cells was retrieved from the NCBI gene bank (Genbank) of the National Center for Bioinformatics in the United States. The gene sequence number is EU660218.1, and the gene fragments ran...

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Abstract

The invention relates to a method for quickly detecting culture medium BHK-21 cell residuals in a foot-and-mouth disease vaccine, and belongs to the field of the biological technology. According to the method, the foot-and-mouth disease vaccine is demulsified through a demulsifying agent, then the total RNA of vaccine residuals is extracted, the Cytb gene of residual cells is detected through the fluorescent quantitative RT-PCR method, meanwhile, specificity analysis is conducted on a target fragment through the PCR-RFLP technology, finally, the content of Cytb in a sample is computed according to a standard curve of a standard substance, and accordingly the aim of quickly and stably detecting the BHK-21 cell residuals in the foot-and-mouth disease vaccine is achieved. The operation process is simple, maneuverability is high, the period is short, sensitivity is high, results are accurate, repeatability is good, and application prospects are wide.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for detecting residual cultured cells in foot-and-mouth disease vaccines, in particular to a method for detecting residual BHK-21 cells in foot-and-mouth disease vaccines by means of real-time fluorescent quantitative PCR. Background technique [0002] Foot-and-mouth disease is an acute, febrile and fast-distance infectious disease carried by cloven-hoofed animals. It infects major livestock species such as pigs, cattle, sheep, and other domestic and wild cloven-hoofed animals. There are more than 70 susceptible animals. kind. The rapid spread and strong infectivity have caused huge economic losses to the animal husbandry industry and seriously affected international trade. Known as the "political and economic disease". The main measures to prevent and control foot-and-mouth disease are to kill diseased animals and immunize susceptible animals with inactivated vaccines of re...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q2545/113C12Q2563/107C12Q2535/138
Inventor 周建国毕峻龙尹革芬段博芳张应国杨贵树
Owner 云南省动物疫病预防控制中心
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