Direct detection of Mycobacterium avium in environmental water and scale samples by loop-mediated isothermal amplification

作者:Nishiuchi Yukiko*; Tamaru Aki; Suzuki Yasuhiko; Kitada Seigo; Maekura Ryoji; Tateishi Yoshitaka; Niki Mamiko; Ogura Hisashi; Matsumoto Sohkichi
来源:Journal of Water and Health, 2014, 12(2): 211-219.
DOI:10.2166/wh.2013.007

摘要

We previously demonstrated the colonization of Mycobacterium avium complex in bathrooms by the conventional culture method. In the present study, we aimed to directly detect M. avium organisms in the environment using loop-mediated isothermal amplification (LAMP), and to demonstrate the efficacy of LAMP by comparing the results with those obtained by culture. Our data showed that LAMP analysis has detection limits of 100 fg DNA/reaction for M. avium. Using an FTA (R) elute card,DNA templates were extracted from environmental samples from bathrooms in the residences of 29 patients with pulmonary M. avium disease. Of the 162 environmental samples examined, 143 (88%) showed identical results by both methods; 20 (12%) and 123 (76%) samples were positive and negative, respectively, for M. avium. Of the remaining 19 samples (12%), seven (5%) and 12 (7%) samples were positive by the LAMP and culture methods, respectively. All samples that contained over 20 colony forming units/primary isolation plate, as measured by the culture method, were also positive by the LAMP method. Our data demonstrate that the combination of the FTA elute card and LAMP can facilitate prompt detection of M. avium in the environment.

  • 出版日期2014