嗜肺军团菌环介导等温扩增检测方法的建立及应用

Establishment and application on detection of Legionella pneumophila using LAMP technology

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 isothermal amplification,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。

Objective To establish a sensitive, rapid, safe and convenient Legionella pneumophila detection methods for applications on port. Methods Legionella pneumophila genome sequence was, analyzed specific gene fragment primers was selected to establish loop-mediated isothermal amplification （loop mediated isothermal amplification LMAP） method, and the reaction condition were optimized. Results Specificity of the established method was good, the sensitivity was 103cfu/ml. The detection capability corresponded to quantitative PCR. Conclusion The Legionella pneumophila LAMP technology detection methods has high sensitivity, specificity, suitable for on-site rapid detection.