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目的:采用Real-time PCR的定量方法对食品中所含蜡样芽胞杆菌的数量进行快速的检测。为食物中毒的快速检测提供有力依据。方法:采用蜡样芽胞杆菌的肠毒素基因(Genebank D17312)作为靶基因,建立PCR反应体系。用已定量的蜡样芽胞杆菌DNA提取液作为外部参照,建立标准曲线进行定量检测。准备模拟样品,把PCR定量方法和国家标准方法的结果进行比较。结果:该反应体系的灵敏度和特异性均能够达到要求。对国标法和PCR定量方法的结果采用T检验分析,t=0.38,v=33,P>0.05,其结果无显著性差异。结论:本实验所采用的Real-time PCR定量方法检测食品中的蜡样芽胞杆菌数量,比国标法操作简单,所需时间短,并有较好的灵敏度和特异性,可以运用到日常工作中。
OBJECTIVE: To rapidly detect the amount of Bacillus cereus contained in food by Real-time PCR quantitative method. Provide a strong basis for the rapid detection of food poisoning. Methods: Bacillus cereus enterotoxin gene (Genebank D17312) as a target gene, the establishment of PCR reaction system. A quantitative Bacillus cereus DNA extract was used as an external reference to establish a standard curve for quantitative detection. Prepare a simulation sample and compare the PCR quantification method with the results of a national standard method. Results: The sensitivity and specificity of the reaction system can meet the requirements. The results of the national standard method and the quantitative PCR method were analyzed by T test, t = 0.38, v = 33, P> 0.05, the results showed no significant difference. Conclusion: The real-time PCR quantitative method used in this study to detect Bacillus cereus in food quantity than the national standard method is simple, the time required is short, and has good sensitivity and specificity, can be applied to daily work .