应用实时荧光PCR快速检测耐甲氧西林金黄葡萄球菌的研究

Rapid detection of methicillin-resistant Staphylococcus aureus by real-time fluorescence PCR

目的应用实时荧光PCR检测耐甲氧西林金黄葡萄球菌(MRSA),为临床快速诊断提供依据。方法以金黄葡萄球菌特异nuc基因及甲氧西林耐药基因mecA基因为靶区域,应用实时荧光PCR检测标本中的金黄葡萄球菌及其耐甲氧西林基因,并与直接培养法比较。结果 69份临床标本中实时荧光PCR方法检出MRSA 41份、MSSA 15份,直接培养法检出MRSA 40份、MSSA 17份,两检查方法的符合率为92.75%。结论对mecA、nuc基因进行实时荧光PCR,能准确、快速地鉴定MRSA。

Objectives To use a real-time PCR assay to detect methicillin--resistant staphylococcus aureus（MRSA）and provide a clinical basis for rapid diagnosis. Methods Primers and probes were designed in accordance with the nuc gene sequences of Staphylococcus aureus and mecA gene sequences of MRSA. Real-time PCR was used to detect Staphylococ- cus aureus and identify the genes of MRSA. The results were compared to a direct culture assay. Results In 69 clinical samples, MRSA was detected by real-time PCR in 41 and MSSA was detected in 15 while MRSA was detected by direct culturing in 40 and MSSA was detected in 17. The agreement of the two assays was 92.75%. Conclusion This study shows that a real-time PCR assay with primers for the mecA and nuc genes is a valuable tool for rapid and precise identifi- cation of MRSA.