新型TaqMan-MGB探针法检测耐甲氧西林金黄色葡萄球菌

目的探讨新型MGB探针在耐甲氧西林金黄色葡萄球菌快速检测中的临床应用价值。方法以TaqMan-MGB探针技术为基础,用已克隆的mecA基因作参比品,实时检测临床标本和阴性参考品。结果所建立方法的最低检测限度为1个基因拷贝/反应,在100~109范围内,CT值(C代表Cycle,T代表阈值)同DNA量的对数呈线性关系;用该方法检测20例MRSA培养阳性的临床标本,结果均为阳性;用该方法检测20个非MRSA细菌株,结果均为阴性。结论所建立的方法适用于临床MRSA快速诊断。

Identification of Meticillin-resistant Staphylococcus aureus by Real-time PCR Based on TaqMan-MGB Probe

OBJECTIVE To develop a real-time PCR assay based on TaqMan-MGB probe for detecting meticillin-resistant Staphylococcus aureus(MRSA). METHODS Primers and MGB probe were chosen in the conserved region of MRSA mecA gene.The efficacy of this assay was evaluated by quantitatively measuring sequential levels of plasmid containing the interest and by detecting clinical specimens and none-MRSA bacterial strains. RESULTS This assay had a detection limit of one genome copy per reaction.A linear standard curve was obtained between 10~0 and 10~9 DNA copies per reaction(r>0.990).No positive results were seen in all non-MRSA bacterial strains tested,while all clinical samples positive for MRSA showed positive results for the mecA gene. CONCLUSIONS These observations suggest that this assay be an excelleot candidate for a standard MRSA detection method.