LAMP技术检测社区获得性非典型肺炎中的嗜肺军团菌感染  

Detection of Legionella pneumophila infection in communityacquired atypical pneumonia by LAMP technique

作　　者：李辉腾 郭旭光[2] 柯茂彬 黎卓华 磨慧玲 吴丽川 李亚东 

机构地区：[1]广东同江医院检验科,广东佛山528300 [2]广州医科大学附属第三医院检验科,广东广州510150

出　　处：《中国医药科学》2017年第23期113-115,149,共4页China Medicine And Pharmacy

基　　金：广东省佛山市医学类科技攻关项目(2015AB002593)

摘　　要：目的探讨环介导恒温扩增技术检测社区获得性肺炎非典型病原体嗜肺军团菌感染的可行性。方法收集195例CAP患者急性期双份合格痰液标本,一份痰液标本用作嗜肺军团菌分离培养,一份痰液标本经处理后-80℃保存至DNA提取运用LAMP技术及荧光定量PCR检测,验证LAMP试验方法的敏感性、特异性及可行性。结果 195例痰培养嗜肺军团菌检出率1.54%(3/195),荧光定量PCR灵敏度达10^(-3)/mL,检出率8.72%(17/195);LAMP方法学灵敏度达10^(-7)/μL,检出率9.23%(18/195);两者阳性符合率94.44%(17/18),两者阴性符合率99.44%(177/178),两者总符合率99.49%(194/195)。结论采用LAMP技术检测临床痰液标本中的嗜肺军团菌具有高度的特异性和敏感性,实验时间约1.5h且操作简便,更合适基层医院开展。Objective To investigate the feasibility of detecting the infection of Legionella pneumophila in the atypical pathogens of community acquired pneumonia by loop mediated constant temperature amplification. Methods Two qualified sputum specimens were collected from 195 patients with CAP in acute stage,one sputum specimen was used for isolation and culture of Legionella pneumophila,one sputum sample was stored at -80℃ for DNA extraction and detected by LAMP technique and fluorescence quantitative PCR.the sensitivity,specificity and feasibility of LAMP test method are verified. Results The detection rate of Legionella pneumophila in 195 cases of sputum culture was 1.54% （3/195）.The sensitivity of fluorescence quantitative PCR was 10-3/mL and the detection rate was 8.72% （17/195）.The sensitivity of the LAMP method was 10-7/L,and the detection rate was 9.23% （18/195）.The positive coincidence rate of both of them was 94.44% （17/18） and the negative coincidence rate of both of them was 99.44% （177/178） and the total coincidence rate of both of the two was 99.49% （194/195）. Conclusion Detection of Legionella pneumophila in clinical sputum samples by LAMP technique has high specificity and sensitivity,the experimental time is about 1.5h and the operation is simple and convenient,which is more suitable for primary hospitals.

关 键 词：LAMP技术 社区获得性非典型肺炎 嗜肺军团菌 痰培养 

分 类 号：R563.1[医药卫生—呼吸系统]