基于QPCR快速检测米饭中产呕吐毒素蜡样芽孢杆菌的研究  

Study on Rapid Detection of Emetic Bacillus Cereus by QPCR

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

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

出　　处：《中国粮油学报》2014年第2期100-104,共5页Journal of the Chinese Cereals and Oils Association

基　　金："十二五"国家科技支撑计划(2011BAK10B06)

摘　　要：为了建立基于QPCR的快速方法检测米饭中产呕吐毒素蜡样芽孢杆菌,首先采用煮沸法提取蜡样芽孢杆菌基因组DNA,并利用普通PCR方法验证引物特异性,然后通过在米饭样品中添加目标菌,模拟受污染的实际样本,用QPCR技术定量检测米饭中产呕吐毒素蜡样芽孢杆菌。结果表明该方法具有快速、特异性强、灵敏度高和稳定性好的优点,能对产呕吐毒素蜡样芽孢杆菌定量。不经过增菌培养,实际样品的检测限为9.8×101CFU/g;经过2 h的增菌培养,检测限能达到100CFU/g;并且米饭中添加其他杂菌后,不影响对蜡样芽孢杆菌的检测。建立的QPCR方法适用于米饭等相关淀粉类食品中产呕吐毒素蜡样芽孢杆菌的检测,从而为监测该菌所致食品污染以及早期相关食物中毒提供快速定量检测方法。Quantitative PCR method has been developed for rapid and accurate detection of emetic Bacillus ce- reus in cooked rice. Experiments were carried out as： DNA was isolated by boiling method and the specificity of primers was tested by conventional PCR. Cooked rice was inoculated with emetic B. cereus for imitating contaminated sample; then was detected by QPCR. The results showed that the QPCR assay was rapid, specific, sensitive and sta- ble, which could detect emetic B. cereus accurately. The limit of detection for emetic B. cereus was 9.8 x 101CFU/g in cooked rice sample ; the assay was suitable for detection of 100 CFU/g emetic B. cereus in cooked rice after two - hour enrichment culture in LB broth. In addition, the Ct value acquired by QPCR assay was maintained even in the presence of other microorganisms which contaminated the cooked rice frequently. The established QPCR method is determined to be efficient, which can be used for the detection of emetic B. cereus in cooked rice as well as other starch - derived food. Therefore, an emergency diagnosis method has been developed for monitoring emetic B. cereu- sus food contamination and detecting early food poison and foodborne diseases.

关 键 词：产呕吐毒素蜡样芽孢杆菌 CESB QPCR 检测 

分 类 号：TS20[轻工技术与工程—食品科学]