耐甲氧西林葡萄球菌荧光定量PCR检测方法的实验研究

目的建立一种简便、特异的mecA基因荧光定量PCR检测方法，用于耐甲氧西林葡萄球菌（MRS）的快速鉴定。方法以煮沸法快速制备DNA模板，采用SYBRGreenI随机参入法，建立mecA基因的实时荧光定量PCR检测体系。并对检测体系的敏感性、特异性和灵敏度进行评价。结果本法对纯菌落的检测敏感性和特异性分别为98．5％和96．9％，检测灵敏度可达10^1CFU／ml，最小检菌量约为3个菌／反应体系。结论本实验所设计的荧光定量PCR方法用于MRS的检测具有快捷、高敏感性、高特异性和高灵敏度的特点。适用于MRS的快速检测。

Rapid Detection of Methcillin-Resistant Staphylococcus with Real Time PCR Assay

Objective To establish a simple real time PCR method for the rapid and accurate identification of Methcillin-resistant Staphylococcus （MRS）. Methods DNA samples were prepared by boiling the bacteria and used as templates for real-time PCR. The primers for real-time PCR were designed based on sequences of the mecA gene of Staphylococcus. 65 MRS strains and 97 non-MRS strains were tested by real-time PCR using SYBR Premix Ex Taq^TM kit. Results The sensitivity and specificity of this detection method were 98.3% and 96.9%, respectively. The minimal detection limits was 3 bacterial cells per reaction or 101 CFU per milliter. Conclusion This is a rapid, accurate and specific method for MRS identification.