实时荧光定量PCR在快速检测耐甲氧西林金黄色葡萄球菌中的应用评估  

Application evaluation of the real-time fluorescence quantitative PCR in the rapid identification of MRSA

作　　者：陈旭[1] 肖淑珍[1] 董丹凤[1] 杨海慧[2] 李生香 倪语星[1] 韩立中[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院临床微生物科,上海200025 [2]上海交通大学医学院附属仁济医院检验科,上海200001 [3]青海西宁市第三人民医院,青海西宁810005

出　　处：《检验医学》2012年第12期1035-1039,共5页Laboratory Medicine

基　　金：上海市自然科学基金资助项目(09ZR1419200);上海市科委择优委托项目(09DZ1907700)

摘　　要：目的评价实时荧光定量聚合酶链反应(PCR)在快速检测耐甲氧西林金黄色葡萄球菌(MRSA)中的应用。方法采用头孢西丁纸片扩散法和mecA基因PCR检测法,将85株临床分离的金黄色葡萄球菌区分为MRSA和甲氧西林敏感金黄色葡萄球菌(MSSA),并采用实时荧光定量PCR对这些菌株进行检测,评价MRSA检测中实时荧光定量PCR与目前常规检测方法的一致性。结果根据头孢西丁纸片扩散法和mecA基因扩增结果进行分组,85株金黄色葡萄球菌中MRSA组菌株45株,MSSA组菌株40株;实时荧光定量PCR检测结果与上述结果完全一致,符合率为100%。结论实时荧光定量PCR检测MRSA的结果与常规方法一致性好,且具有操作简便、耗时短等特点。作为一种快速检测方法,实时荧光定量PCR能将金黄色葡萄球菌的鉴定和MRSA的筛选结合起来,对于指导临床用药及MRSA医院感染控制具有重要意义。Objective To evaluate the application significance of the real-time fluorescence quantitative polymerase chain reaction （PCR） in the rapid identification of methicillin-resistant Staphylococcus aureus （MRSA）. Methods A total of 85 Staphylococcus aureus strains were isolated from clinical samples, and MRSA and methicillin-sensitive Staphylococcus aureus （MSSA） were detected and differentiated by cefoxitin disk diffusion method and conventional PCR amplification of mecA gene. These strains were also detected by the real-time fluorescence quantitative PCR. The accordance of the real-time fluorescence quantitative PCR with conventional methods was evaluated. Results According to the results of the cefoxitin disk diffusion method and conventional PCR amplification of mecA gene, 45 out of 85 strains were MRSA, and 40 out of 85 strains were MSSA. The real-time fluorescence quantitative PCR had an excellent accordance （ 100% ） with those methods. Conclusions The real-time fluorescence quantitative PCR has an excellent accordance with conventional methods for detecting MRSA, and it has the advantages of ease of performance and costing short turn-around time. As a rapid identification method, the real-time fluorescence quantitative PCR can identify MRSA accurately, which is helpful for the clinical therapy of MRSA infections and for the control of MRSA transmission in hospitals.

关 键 词：金黄色葡萄球菌 耐甲氧西林金黄色葡萄球菌 实时荧光定量聚合酶链反应 快速检测 

分 类 号：R446.5[医药卫生—诊断学]