Method for rapidly detecting and parting staphylococcal enterotoxin in food by fluorescent polymerase chain reaction (PCR) method

A technology of Staphylococcus enterica and Staphylococcus, which is applied in the field of molecular detection of microbial toxins, can solve the problems of small detection throughput, great health hazards for operators, and long detection time, and achieve improved typing speed, safety advantages, and detection The effect of flux enhancement

Inactive Publication Date: 2011-05-18
NANJING INST OF PROD QUALITY INSPECTION
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Problems solved by technology

For example, the invention patent with the application number 200910071033.4 "Detection method of staphylococcal enterotoxin genotype and genotype detection method" adopts the above method, and its disadvantages include: long detection time, PCR amplification and electrophoresis detection will take more than 3 hours; The detection throughput is small, and each elec

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  • Method for rapidly detecting and parting staphylococcal enterotoxin in food by fluorescent polymerase chain reaction (PCR) method
  • Method for rapidly detecting and parting staphylococcal enterotoxin in food by fluorescent polymerase chain reaction (PCR) method
  • Method for rapidly detecting and parting staphylococcal enterotoxin in food by fluorescent polymerase chain reaction (PCR) method

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Embodiment Construction

[0020] 1. Experimental materials, reagents and instruments

[0021] Food total DNA extraction kit, China TIANGEN company; SYBR Green fluorescent quantitative PCR reagent buffer solution, Japan Takara company; primers are entrusted to Shanghai Sangong company to synthesize. Fast fluorescent quantitative PCR instrument, ABI 7500 Fast, American ABI Company; high-speed centrifuge, Sigma3-18K, American Sigma Company; slap homogenizer, China Liuyi Company; UV spectrophotometer, American Perkin Company; mini VIDAS enzyme Fluorescence Immunoassay Analyzer, BioMérieux, France.

[0022] 2. Collection of food samples contaminated with Staphylococcus aureus

[0023]Taking advantage of the advantages of the National Food Quality Inspection Center, in the food supervision and random inspection and entrusted inspection, according to the traditional process of selective culture and plasma coagulase biochemical identification in the national standard GB4789, collect food samples positive for ...

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Abstract

The invention belongs to the field of molecular detection for microbial toxin in food science and technology, in particular to rapid detection and parting of 11 serotypes of staphylococcal enterotoxin in food by applying the flows such as total DNA component extraction and SYBRGreen fluorescent quantitative PCR (polymerase chain reaction) rapid detection. Results show that the staphylococcal enterotoxin in food can be effectively typed according to the situation whether marked amplification curves appear in 35 PCR cycles by utilizing the method, thus the method has the characteristics of specificity aiming at enterotoxin genes of different serotypes, sensitivity, accuracy, rapidness and convenience and the like; and compared with the existing method, the method provided by the invention has the advantages of greatly improving the detection and parting efficiency and providing a new way for risk analysis of food pollution caused by the staphylococcal enterotoxin of different types.

Description

technical field [0001] The invention belongs to the field of molecular detection of microbial toxins in food science and technology, and specifically relates to a rapid high-throughput detection and typing method for 11 serotype genes of staphylococcal enterotoxins in food by using a fluorescent quantitative PCR method and an application thereof. Background technique [0002] Staphylococcus aureus ( Staphylococcus aureus , Staphylococcus aureus) is one of the most common foodborne pathogens, which not only cause a large number of cases of food poisoning and purulent infection, but also can cause explosive epidemics. The virulence factors of Staphylococcus aureus include various proteins produced during invasion, such as thermostable nuclease, hyaluronidase, lipase, hemolysin and enterotoxin. Among them, staphylococcal enterotoxin (Staphylococcal Enterotoxin, SE) is the key pathogenic factor that causes human food poisoning and septic shock, and its essence is to pass T cell...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张驰杨军凌睿刘新梅周骏贵
Owner NANJING INST OF PROD QUALITY INSPECTION
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