基于不同靶基因的荧光定量PCR快速检测蜡样芽孢杆菌的研究

Development of real-time PCR method based on different target genes for rapid detection of Bacillus cereus

摘要: 为了建立一种基于不同靶基因的快速灵敏检测食品中产与不产呕吐毒素蜡样芽孢杆菌的荧光定量PCR方法,本研究采用煮沸法提取基因组DNA,用普通PCR方法验证引物的特异性,通过在米饭中添加不同浓度的产呕吐毒素蜡样芽孢杆菌模拟受污染的食品,采用本研究构建的荧光定量PCR方法对米饭进行检测。结果表明基于不同靶基因设计的两对引物具有特异性强和扩增效率高等优点,荧光定量PCR方法能对污染米饭中蜡样芽孢杆菌准确定量,检测限能达到101CFU/g。建立的荧光定量PCR方法特异、灵敏和准确,适用于食品中蜡样芽孢杆菌的快速定量检测。

Abstract: To develop a rapid and sensitive real-time PCR method to differentiate emetic Bacillus cereus and non- emetic B.cereus based on different target genes.DNA was isolated by boiling method and used to test the specificity of primers by PCR.The rice was inoculated by emetic B.cereus for imitating contaminated food and detected by real- time PCR method.The results showed that primers were specific and high of amplification efficiency, the B.cereus could be quantified accurately by real-time PCR and the limit of detection was 101CFU /g in the contaminated rice.The novel developed real- time PCR method was specific, sensitive and accurate, which could be used for the rapid detection of B.cereus in food.