结核分枝杆菌LAMP检测方法的建立及应用  

Establishment and application of LAMP for Mycobacterium tuberculosis

作　　者：陈钰 罗力涵 李莎 樊小刚 裴程程 王嫱 张波 CHEN Yu LUO Li-han LI Sha FAN Xiao-gang PEI Cheng-cheng WANG Qiang ZHANG Bo(Shenyang International Travel Healthcare Center, Shenyang, Liaoning 110016, China)

机构地区：[1]沈阳国际旅行卫生保健中心,辽宁沈阳110016

出　　处：《中国国境卫生检疫杂志》2017年第4期242-247,共6页Chinese Journal of Frontier Health and Quarantine

基　　金：国家质检总局科技计划项目(2015IK170)

摘　　要：目的建立一种基于环介导等温扩增(LAMP)的结核分枝杆菌核酸快速检测方法,并应用于出入境人群的肺结核样本检测。方法针对结核分枝杆菌插入序列6110(IS6110)设计5组LAMP引物,利用LAMP实时浊度仪筛选出1组最佳引物,用20株临床致病菌株进行特异性检测,用10倍梯度稀释法进行灵敏度测定,用25份痰培养阳性样本对阳性率进行评估。结果筛选出1组最佳引物TB159,该引物特异性良好,与20株常见致病菌株无交叉反应。此方法的最低检测限为24.77 pg/μl,对25份痰培养阳性样本检测的阳性率为100%。结论建立了一种特异性好、灵敏度高的LAMP快速检测方法,可以应用于结核病检测。Objective To establish a rapid detection method for Mycobacterium tuberculosis nucleic acid based on Loop-mediated isothermal amplification（LAMP）,and apply it to tuberculosis diagnosis in the entry-exit population.Methods Five pairs of LAMP primer were designed. A set of best primer was selected by using LAMP Real-time turbidity meter,20 bacteria strains were used to test specificity,the 10 times gradient dilution method was used to determine the sensitivity,25 sputum culture positive samples was used to evaluate positive rate. Results A set of best primer TB159 was selected,there was no cross reaction with 20 common bacterial strains. The detection limit was 24.77 pg/μl,the positive rate was 100% in testing 25 sputum culture positive samples. Conclusion A high specificity and sensitivity LAMP rapid detection method was established,which could be applied to tuberculosis diagnosis.

关 键 词：环介导等温扩增技术 结核分枝杆菌 插入序列6110 

分 类 号：R392.1[医药卫生—免疫学]