实时荧光PCR快速检测耐甲氧西林金黄色葡萄球菌的临床评估  

Real-time fluorescence PCR for detection of methicillin-resistant Staphylococcus aureus

作　　者：李耀华[1] 费春荣[2] 叶爱青[2] 

机构地区：[1]国家食品药品监督管理局医疗器械技术审评中心,北京100044 [2]浙江大学医学院附属邵逸夫医院检验科,浙江杭州310016

出　　处：《诊断学理论与实践》2011年第5期449-453,共5页Journal of Diagnostics Concepts & Practice

摘　　要：目的:采用实时荧光PCR检测耐甲氧西林金黄色葡萄球菌(MRSA),并评价该方法在临床上的应用价值。方法:将临床分离的62株金黄色葡萄球菌(金葡菌)和35株非金黄色葡萄球菌同时采用分离培养及药物敏感试验(药敏试验)和实时荧光PCR法进行检测并比较,对2种方法检测结果不符的菌株辅以测序法进行最后鉴定,并确定实时荧光PCR法的检测灵敏度。结果:分离培养法与实时荧光PCR法对金葡菌检测的一致率为100%,都检测出了62株金葡菌阳性株。实时荧光PCR法与分离培养药敏试验法MRSA检测的符合率为96.77%,而分离培养药敏试验法检测为阴性而实时荧光PCR法检测为阳性的2株待检菌的测序结果显示为MRSA阳性株。最后确定实时荧光PCR法检测MRSA的临床灵敏度为1×103 cfu/mL。结论:实时荧光PCR法可用于临床对MRSA的快速检测,具有较高的临床价值。Objective Using real-time fluorescence PCR for the detection of methicillin-resistant Staphylococcus aureus（MRSA） and to assess it clinical value.Methods Real-time fluorescence PCR and isolation-culture-K-B paper disk diffusion method were performed to analyze 62 Staphylococcus aureus strains and 35 non-Staphylococcus aureus strains isolated clinically.Samples with different testing results by these two methods were further examined by sequencing method for final identification.Results Sixty-two Staphylococcus aureus were identified by culture method and real-time fluorescence PCR method.Of the 62 clinical isolated Staphylococcus aureus 41 isolates were defined as MRSA by K-B paper disk diffusion method,and 43 isolates were defined as MRSA by real-time fluorescence PCR.The two isolates defined as positive by real-time fluorescence PCR and negative by K-B paper disk diffusion method were proved to be MRSA positive by sequencing method.The sensitivity of PCR for detecting MRSA was 1×103 cfu/mL.Conclusions Real-time fluorescence PCR could be used clinically for the rapid detection of MRSA with high clinical value.

关 键 词：耐甲氧西林金黄色葡萄球菌 细菌培养 实时荧光聚合酶链反应 

分 类 号：R466.1[医药卫生—临床医学]