实时荧光定量聚合酶链反应快速检测下呼吸道痰液标本耐甲氧西林金黄色葡萄球菌的临床应用  

Real-time fluorescence quantitation polymerase chain reaction for the detection of methicillin-resistant Staphylococcus aureus from lower respiratory tract

作　　者：周俊[1] 陈旭[2] 李敏[1] 韩立中[2] 杜娟[1] 杨昆[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院呼吸内科,上海200025 [2]上海交通大学医学院附属瑞金医院临床微生物科,上海200025

出　　处：《上海医学》2013年第1期23-26,共4页Shanghai Medical Journal

基　　金：上海市医学会临床医学科研专项资金资助项目(SMJ2010002)

摘　　要：目的应用实时荧光定量聚合酶链反应(PCR)检测耐甲氧西林金黄色葡萄球菌(MRSA)核酸,寻求呼吸道标本MRSA的快速检测技术。方法 收集临床分离的75株金黄色葡萄球菌菌株及97份下呼吸道感染患者的合格痰液标本,应用实时荧光定量PCR快速检测法检测标本中Nuc及MecA基因,并以常规培养+头孢西丁纸片扩散法的检测结果为标准,判断实时荧光定量PCR与传统方法检测结果的一致性,及该方法对下呼吸道痰液标本检测的敏感度、特异度。结果以常规培养+头孢西丁纸片扩散法的检测结果为对照,实时荧光定量PCR快速检测法检测金黄色葡萄球菌菌株中MRSA的敏感度及特异度均为100.0%,检测下呼吸道痰液标本MRSA的敏感度为100.0%、特异度为84.7%。结论应用实时荧光定量PCR快速检测下呼吸道痰液标本MRSA的敏感度高,特异性强,操作时间短,与常规培养+药物敏感试验的检测结果具有良好的一致性,具有较高的临床应用价值。Objective To measure the nucleic acid of methicillin-resistant Staphylococcus aureus （MRSA） by real-time fluorescence quantitation polymerase chain reaction （PCR）, so as to find a rapid detection method for the MRSA from airway specimens. Methods Totally 75 strains of staphylococcus aureus and 97 sputum specimens of lower respiratory tract were collected. Real-time fluorescence quantitation PCR was used to detect Nun and MecA genes, and its sensitivity and specificity were compared with isolation-culture-K-B diffusion method . Results Compared with isolation-culture-K-B diffusion method, the real-time fluorescence quantitive PCR had 100.0% sensitivity and 100.0% specificity for the 75 strains of staphylococcus aureus and had 100.0% sensitivity and 84.7% specificity for the 97 samples of lower respiratory tract. Conclusion Real-time fluorescent quantitation PCR has high sensitivity, specificity and short operating time in detecting MRSA from clinical specimens of lower respiratory tract. The POR result is consistent with that of the conventional culture plus susceptibility method.

关 键 词：耐甲氧西林金黄色葡萄球菌 下呼吸道标本 细菌培养 实时荧光定量聚合酶连反应 快速检测 

分 类 号：R440[医药卫生—诊断学]