Detection of Clostridium perfringens α-toxin and Staphylococcus aureus Enterotoxin A in Dairy with Double PCR Technique

The study was to establish a duplicate PCR detection method for Clostridium perfringens α-toxin and Staphylococcus aureus enterotoxin A.in milk and to guarantee the safety of dairy products.According to gene sequences of Clostridium perfringens α-toxin and Staphylococcus aureus enterotoxin A,two pairs of specific primers were designed.The DNA templates were extracted respectively from the cultured bacterial liquids of Clostridium perfringens and Staphylococcus aureus and used for DNA amplification.The specificity and sensibility of the method were also detected,and some clinical samples were checked in this way.The result showeds that,based on the optimized reaction conditions of PCR,samples were processed DNA amplification for Clostridium perfringens and Staphylococcus aureus.The matching prospective bands were obtained in both bacteria.But this reaction system could not amplify products in other bacterium like Bacillus coli,which indicated this method had specificity.Even for the 1000-times-diluted bacterial liquids of Clostridium perfringens and Staphylococcus aureus,this method could also amplify the target gene fragment,which meant its high sensitive.This PCR method was also compared with routine bacteriology culture method,and the coincidence rate of these two methods was above 90%.In conclusion,a duplicate PCR detection method for Clostridium perfringens α-toxin and Staphylococcus aureus enterotoxin A had been established and it had high sensitivity and specificity.