应用双重PCR快速检测耐甲氧西林金黄色葡萄球菌

Application of duplex PCR for rapid detection of methicillinresistant Staphylococcus aureus

摘要:

目的应用双重聚合酶链反应（PCR）技术，建立金黄色葡萄球菌及耐甲氧西林金黄色葡萄球菌（MRSA）的快速检测方法，指导临床及时、合理使用抗菌药物，防止MRSA的扩散。方法 根据金黄色葡萄球菌血浆凝固酶基因Coag和耐药基因mecA设计引物，调整PCR扩增反应各参数，建立快速、准确扩增Coag和mecA基因的双重PCR体系；应用双重PCR对临床分离鉴定的85株金黄色葡萄球菌同时扩增Coag和mecA基因片段，并将PCR扩增结果与苯唑西林-高盐琼脂筛选试验（OSAS）的结果进行比对。结果生化常规试验分离鉴定85株金黄色葡萄球菌，通过OSAS试验，共检出MRSA 53株，MRSA检出率为62.35%。双重PCR快速检测MRSA，85株金黄色葡萄球菌均扩增出Coag基因片段，其中53株扩增出mecA基因片段。双重PCR检测MRSA的结果与生化常规试验分离鉴定MRSA结果一致。结论双重PCR法能快速、同时检测金黄色葡萄球菌血浆凝固酶Coag基因和耐药基因mecA，有助于及早检出MRSA，指导临床合理用药，控制MRSA传播。

Abstract:

ObjectiveTo establish duplex polymerase chain reaction(PCR) system for fast detecting Staphylococcus aureus (S. aureus) and methicillinresistant S. aureus（MRSA）, guiding rational use of antimicrobials in clinical practice, and preventing MRSA spreading.MethodsTwo pairs of primers according to coagulase gene (Coag) and drug resistance gene (mecA) of S. aureus were designed, duplex PCR system was established and applied for amplifying Coag and mecA in 85 clinical S.aureus strains, amplification reaction result was compared with that of oxacillinsalt agar screening （OSAS） test.ResultsOf 85 clinical S.aureus isolates, 53 (62.35%) were MRSA detected by OSAS test. All 85 S.aureus isolates were detected Coag gene fragments by duplex PCR， and 53 of which were also detected mecA gene fragments, PCR results were consistent with OSAS test result for detecting MRSA.ConclusionDuplex PCR can detect Coag and mecA of S.aureus rapidly and simultaneously, and is helpful for early detection of MRSA, guidance of rational use of antimicrobial agents,and control of MRSA transmission.