实时荧光PCR定量检测食品中单增李斯特菌

目的建立快速、敏感、特异的食品中单增李斯特菌检测方法。方法针对单增李斯特菌溶素A基因（hlyA）设计一对引物和一条探针，并用该引物和探针运用实时荧光PCR技术对单增李斯特菌的DNA、细胞、质粒和样品进行实时荧光PCR定量检测。结果利用实时荧光PCR技术，建立了DNA校正曲线、细胞校正曲线和质粒校正曲线。DNA校正曲线在1～32CFU/ml、细胞校正曲线在32—320CFU／ml、质粒校正曲线在1—37Copies／ml，线形关系良好，且三种校正曲线检测样品得出的结果基本吻合。结论本试验建立起来的实时荧光PCR定量检测单增李斯特菌的方法灵敏度高、特异性好、准确，可应用于食品中单增李斯特菌的检测。

Development of Real-time PCR Method for Quatitaive Detection of Listeria monocytogenes in Foods

Objective To establish a rapid, sensitive and specific method for detection of Listeria monocytogenes(Lm) in the import and export foods.Method A pair of oligonucleotide primers and a probe were designed with listeriolysion A (hlyA) gene as target sequence DNA, cell, plasmid of Lm were amplified by real-time PCR technique.Results This trial established DNA standard curve, cell standard curve and plasmid standard curve by real-time PCR technique. The results obtained by the three standard curves were mainly the same.Conclusion Real-time PCR can be used as a sensitive, specific and accurate method for detection of Lm. The inspection system developed in this experiment can be used to do Listeria monocytogenes inspection work in the department of inspection and quarantine.