荧光定量PCR检测细菌NDM-1基因方法的探讨  

Application of real-time fluorescent quantitative PCR in NDM-1 detection of bacteria

作　　者：张建明[1] 黄吉城[2] 吴健[3] 王向阳[1] 何洪涛[1] 陈姗[3] 师永霞[2] 李小波[2] 洪烨[2] 

机构地区：[1]广东国际旅行卫生保健中心,广州510635 [2]广东出入境检验检疫局技术中心卫生检疫实验室,广州510700 [3]广东国际旅行卫生保健中心检验科,广州510635

出　　处：《中国卫生检验杂志》2013年第1期133-135,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的:将Taqman荧光定量PCR技术与SYBRGreen染料RT-PCR技术比较后,应用于检测细菌NDM-1基因。方法:使用Taqman荧光定量PCR方法与SYBRGreen染料RT-PCR对已知特性的标准株中NDM-1基因检测,对比其扩增灵敏度。结果:Taqman荧光定量PCR技术灵敏度高于SYBRGreen染料RT-PCR法,所测标准株中NDM-1基因转录水平可高达7.5×104拷贝/μl,比SYBRGreen染料RT-PCR技术检测灵敏度高7.8倍。结论:Taqman荧光PCR技术对超级耐药菌NDM-1基因具有较高的敏感性和准确性,可以作为对超级细菌进行监控与防控的方法。Objective：In this study,TaqmanRT-PCR and SYBRGreenRT-PCR were applied for quantification of NDM-1 gene in bacteria.Methods： After detection of NDM-1 gene with Taqman RT-PCR and SYBRGreenRT-PCR,the sensititity was compared.Results： TaqmanRT-PCR was more sensitive and the resultant concentration of RNA level was up to 7.5×104 copies/μl.Compared with SYBR Green RT-PCR,it was up to 7.8-fold higher when determined by TaqMan RT-PCR.Conclusion： The detection of NDM-1 gene in bacteria with TaqmanRT-PCR was proved to be more sensitive and accurate,so Taqman RT-PCR could be suggested as a way to prevent and control super bacteria.

关 键 词：NDM-1 Taqman荧光定量PCR SYBRGreen染料RT-PCR 

分 类 号：R446.5[医药卫生—诊断学]