李斯特菌属荧光定量PCR检测方法的建立  

The establishment of detection method of the Listeria using fluorescence quantitative PCR

作　　者：杨秀娟[1] 张文玲[1] 徐惠芳[1] 田茵[1] 赵嘉珩[1] 李勐伟 吴燕[1] 汪琳[1] 郭敏卓[1] 

机构地区：[1]北京国际旅行卫生保健中心,北京100088

出　　处：《中国国境卫生检疫杂志》2015年第1期6-12,共7页Chinese Journal of Frontier Health and Quarantine

基　　金：国家质检总局科技计划项目(2013IK220)

摘　　要：目的采用荧光定量PCR技术,建立可以同步检测李斯特菌属与单增李斯特菌的快速检测方法。方法选取单增李斯特菌hlyM基因和李斯特菌属23Sr DNA基因为靶基因,分别设计引物和探针,在构建二者的阳性重组质粒基础上,对李斯特菌属和单增李斯特菌进行荧光定量PCR的检测。结果建立的李斯特菌属和单增李斯特菌单重荧光定量PCR检测方法与双重荧光定量PCR的检测方法敏感度一致,分别为18.6拷贝/μl和23.2拷贝/μl。结论建立的双重荧光定量PCR方法可以同步检测李斯特菌属与单增李斯特菌。Objective To establish a duplex real-time PCR-based assay for the simultaneous quantitative detection of Listeria spp and the L. monocytogenes. Methods The hlyM（L. monocytogenes）and 23 SrDNA（Listeria spp）genes were selected as the target genes, the primers and probes were designed, and reaction condition had been optimized. Results The real-time PCR assay to detect Listeria spp and the L. monocytogenes had been established.e The sensitivity of duplex detection of Listeria spp and L. monocytogenes was 18.6 copies/μl and 23.2 copies/μl,respectively, it was the same with that of the single detection method. Conclusion The established duplex real-time PCR assay can detect Listeria monocytogenes and Listeria spp.

关 键 词：单增李斯特菌 李斯特菌 双重荧光PCR 

分 类 号：R183.35[医药卫生—流行病学]