首页 | 本学科首页   官方微博 | 高级检索  
   检索      


Application of EMA-qPCR as a complementary tool for the detection and monitoring of <Emphasis Type="Italic">Legionella</Emphasis> in different water systems
Authors:Tian Qin  Zhengan Tian  Hongyu Ren  Guangchun Hu  Haijian Zhou  Jinxing Lu  Chengwang Luo  Zunyu Liu  Zhujun Shao
Institution:(1) State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Changbai Road, Changping, Beijing, 102206, People’s Republic of China;(2) Shanghai Entry-Exit Inspection and Quarantine Bureau, No. 1208, Minsheng Road, Pudong New Area, Shanghai, 200135, People’s Republic of China;(3) Jinan Municipal Center for Disease Control and Prevention, No. 2, Weiliu Road, Jinan, 250022, People’s Republic of China;
Abstract:Legionella are prevalent in human-made water systems and cause legionellosis in humans. Conventional culturing and polymerase chain reaction (PCR) techniques are not sufficiently accurate for the quantitative analysis of live Legionella bacteria in water samples because of the presence of viable but nonculturable cells and dead cells. Here, we report a rapid detection method for viable Legionella that combines ethidium monoazide (EMA) with quantitative real-time PCR (qPCR) and apply this method to detect Legionella in a large number of water samples from different sources. Results yielded that samples treated with 5 μg/ml EMA for 10 min and subsequently exposed to light irradiation for 5 min were optimal for detecting Legionella. EMA treatment before qPCR could block the signal from approximately 4 log10 of dead cells. When investigating environmental water samples, the percent-positive rate obtained by EMA-qPCR was significantly higher than conventional PCR and culture methods, and slightly lower than qPCR. The bacterial count of Legionella determined by EMA-qPCR were mostly greater than those determined by culture assays and lower than those determined by qPCR. Acceptable correlations were found between the EMA-qPCR and qPCR results for cooling towers, piped water and hot spring water samples (r = 0.849, P < 0.001) and also found between the EMA-qPCR and culture results for hot spring water samples (r = 0.698, P < 0.001). The results indicate that EMA-qPCR could be used as a complementary tool for the detection and monitoring of Legionella in water systems, especially in hot spring water samples.
Keywords:
本文献已被 PubMed SpringerLink 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号