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