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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 rapid detection method for simultaneous detection of Listeria and Listeria monocytogenes using fluorescence quantitative PCR. Methods The genes of hlyM gene of Listeria monocytogenes and 23Sr gene of Listeria were selected as target genes. Primers and probes were designed respectively. Based on the constructed positive recombinant plasmids of both Listeria and Listeria monocytogenes Fluorescent quantitative PCR detection. Results The results of single fluorescence quantitative PCR assay of Listeria monocytogenes and Listeria monocytogenes were identical with that of double fluorescence quantitative PCR, which were 18.6 copies / μl and 23.2 copies / μl, respectively. Conclusion The established double fluorescence quantitative PCR method can simultaneously detect Listeria and Listeria monocytogenes.