三重荧光PCR技术在食品产气荚膜梭菌毒素基因检测中的应用
Application of Triple Fluorescent PCR Technique in the Detection of Food Toxin Genes of Clostridium perfringens
邹淑慧 1何震 1唐美秀1
作者信息
摘要
目的:建立食品中产气荚膜梭菌常见 3 种毒素(α、β、ε)的基因cpa、cpb、ext的三重荧光PCR检测方法.方法:设计产气荚膜梭菌的α、β、ε 3 种毒素基因的引物和探针,优化退火温度,建立三重荧光PCR方法,探讨其特异性、灵敏度.结果:该方法的特异性较强,仅能检测出产气荚膜梭菌,对其他梭菌与常见食源性致病菌的扩增结果均为阴性;三重荧光PCR体系的检测灵敏度达到 0.000 1 ng·μL-1.用建立的方法对实验室样品进行鉴定,发现 2 份食品样品中有产气荚膜梭菌(均为A型).结论:本研究建立的三重荧光PCR方法的特异性、灵敏度均较高,且操作简单、检测速度快,适用于食品中产气荚膜梭菌的检测.
Abstract
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.
关键词
三重荧光PCR/产气荚膜梭菌/毒素Key words
triple fluorescence PCR/Clostridium perfringens/toxin引用本文复制引用
出版年
2024
NETL
NSTL
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