嗜肺军团菌环介导等温扩增检测方法的建立及应用

张晓航1，陆晔1，周娴1，周启明1，李俊1，田桢干1，章琪1，吴凡2，杜正平2，方筠1

　　1.上海国际旅行卫生保健中心，上海 200335；2.广州华峰生物科技有限公司

　　摘要：目的 建立一种灵敏、快速、安全、方便适合口岸现场应用的嗜肺军团菌检测方法。方法分析嗜肺军团菌基因组序列，选择特异性基因片段设计引物，建立环介导等温扩增（loop mediated isothermalamplification， LMAP）方法，并优化反应条件。结果建立的方法特异性好，灵敏度可达103cfu/ml，检测能力与荧光定量PCR法相当。结论应用环介导恒温扩增技术建立的嗜肺军团菌检测方法，灵敏度高，特异性好，适合现场快速检测。

　　关键词：嗜肺军团菌；环介导等温扩增法；检测；应用

　　中图分类号：R392.1　文献标识码：B

　　Establishment and application on detection of Legionellapneumophila

　　using LAMP technology

　　ZHANG Xiao-hang*， LU Ye, ZHOU Xian, ZHOU Qi-ming, LI Jun, TIANZhen-gan,

　　ZHANG Qi, WU Fan, DU Zheng-ping, FANG Yun

　　*Shanghai International Travel Healthcare Center, Shanghai200335, China

　　Abstract:   Objective   To establish a sensitive,rapid, safe and convenient Legionella pneumophila detection methodsfor applications on port.   Methods   Legionellapneumophila genome sequence was, analyzed specific gene fragmentprimers was selected to establish loop-mediated isothermalamplification (loop mediated isothermal amplification LMAP) method,and the reaction condition were optimized.   Results  Specificity of the established method was good, the sensitivity was103cfu/ml. The detection capability corresponded to quantitativePCR.   Conclusion   The Legionella pneumophila LAMPtechnology detection methods has high sensitivity, specificity,suitable for on-site rapid detection.

　　Key words:   Legionella pneumophila; loop-mediatedisothermal amplification (LAMP); Detect; Application

　　《中国国境卫生检疫杂志》2014年2月刊