Application of Triple Fluorescent PCR Technique in the Detection of Food Toxin Genes of Clostridium perfringens
Objective:To establish a triple fluorescence PCR method for the detection of three common toxins(α,β and ε)of Clostridium perfringens in food.Method:Primers and probes for three toxin genes of Clostridium perfringens,α,β and ε were designed,the annealing temperature was optimized,and a triple fluorescence PCR method was established to explore its specificity and sensitivity.Result:The specificity of this method was strong,and it could only detect Clostridium perfringens,and the amplification results of other Clostridium perfringens were negative,and the detection sensitivity of the triple fluorescence PCR system reached 0.000 1 ng·μL-1.The laboratory samples were identified by the established method,and Clostridium perfringens(both type A)was found in 2 food samples.Conclusion:The triple fluorescence PCR method established in this study has high specificity,sensitivity,simple operation and fast detection speed,and is suitable for the detection of Clostridium perfringens in food.
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