环介导恒温扩增法检测肉制品中致病微生物

目的:建立一种食品中微生物核酸快速检测技术,在此基础上开发的简便、快速、灵敏,不依赖仪器适合基层应用的核酸快速检测试剂盒;并探索该试剂盒在肉及肉制品微生物检测中的应用。方法:根据单核细胞增生李斯特氏菌特有的靶序列vir R基因设计引物,采用环介导恒温扩增技术构建检测体系,优化、验证检测的特异度、检出限等指标,并与传统分离培养法比较。结果:本研究中的检测方法及试剂盒能专一的检出肉制品中的单增李斯特菌,对其他近源菌株检测呈阴性结果,检出限达1.81×103CFU/mL,与传统方法具有相同的灵敏度;检测操作简便快速,极少的设备依赖,适合规模推广。

Rapid Detection of Listeria monocytogenes in Meat Product Using Loop-Mediated Isothermal Amplification Method

Objectives： To establish a method for rapid detection of microbial nucleic acids in food, develop a convenient, rapid and sensitive nucleic acid detection kit, and apply the kit for Listeria monocytogenes detection in meat product. Methods： According to the sequence of the characteristic target gene vir R, specific primer pairs were designed. A detection system was constructed using loop-mediated isothermal amplification and optimized. The established method was analyzed for its specificity and limit of detection, and compared with the conventional methods （based on isolation and subsequent culture）. Results： The detection method and kit developed in this study was specific to Listeria monocytogenes detection in meat product and presented negative results for other related species. The limit of detection was 1.81 × 10^3 CFU/mL, and the sensitivity was consistent with that of the conventional methods. Other advantages such as simple and rapid detection and few needs for instruments were also achieved, thereby deserving to be popularized.