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

作　　者：张志鸿[1,2] 甘蓓 谭强来[1] 王力均[1] 刘成伟[4] 游兴勇[4] 许恒毅[1] 魏华[2] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]南昌大学中德联合研究院,江西南昌330047 [3]江西省产品质量监督检测院,江西南昌330029 [4]江西省疾病预防控制中心,江西南昌330000

出　　处：《食品工业科技》2013年第23期160-163,共4页Science and Technology of Food Industry

基　　金：食物中毒因子解析技术研究(2011BAK10B06);"十二五"国家科技支撑计划项目;基于纳米荧光材料标记的免疫层析检测技术的定量方法学建立(20114BAB214017);江西省自然科学基金

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。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 10^1 CFU/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.

关 键 词：蜡样芽孢杆菌 CESB GYRB 荧光定量PCR 检测 

分 类 号：TS201.6[轻工技术与工程—食品科学]