Lu Qin-feng, Zheng Wei, Luo Peng, Wu Zhong-hua, Li He, Shen Jian-gen
College of Medicine, Zhejiang University, Hangzhou 310058, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2010 May;39(3):305-10. doi: 10.3785/j.issn.1008-9292.2010.03.015.
To establish a simple and rapid molecular detection for Legionella pneumophila.
The loop-mediated isothermal amplification (LAMP) was applied for detection of Legionella pneumophila. A set of primers were designed to identify six special areas in mip gene of Legionella pneumophila. Genomic DNAs from 13 bacterial strains,including 8 Legionella pneumophila strains and 5 other bacterial strains were amplified by LAMP and general PCR method to evaluate the specificity and sensibility of LAMP.
All positive tubes produced visible white precipitation, and no precipitation was observed in others. By adding smart green fluorescent dye, all Legionella pneumophila positive tubes presented a strong green fluorescence, while others showed weak fluorescence. The detection rate of LAMP was higher than that of general PCR. The detection limits were 576fg with genomic DNA of Legionella pneumophila,and 8 cfu/mL with positive water samples.
LAMP detection of Legionella pneumophila is an effective and low-cost method with high specificity and sensitivity requiring no special equipment.
建立一种简单快速的嗜肺军团菌分子检测方法。
应用环介导等温扩增技术（LAMP）检测嗜肺军团菌。设计一组引物以识别嗜肺军团菌mip基因中的六个特定区域。采用LAMP和常规PCR方法对13株细菌（包括8株嗜肺军团菌菌株和5株其他细菌菌株）的基因组DNA进行扩增，以评估LAMP的特异性和敏感性。
所有阳性管均产生可见的白色沉淀，其他管未观察到沉淀。加入智能绿色荧光染料后，所有嗜肺军团菌阳性管呈现强烈的绿色荧光，而其他管显示弱荧光。LAMP的检测率高于常规PCR。嗜肺军团菌基因组DNA的检测限为576fg，阳性水样的检测限为8 cfu/mL。
LAMP检测嗜肺军团菌是一种有效且低成本的方法，具有高特异性和敏感性，无需特殊设备。
