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Direct detection of Mycobacterium avium in environmental water and scale samples by loop-mediated isothermal amplification
Author(s) -
Yukiko Nishiuchi,
Aki Tamaru,
Yasuhiko Suzuki,
Seigo Kitada,
Ryoji Maekura,
Yoshitaka Tateishi,
Mamiko Niki,
Hisashi Ogura,
Sohkichi Matsumoto
Publication year - 2013
Publication title -
journal of water and health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.482
H-Index - 59
eISSN - 1996-7829
pISSN - 1477-8920
DOI - 10.2166/wh.2013.007
Subject(s) - loop mediated isothermal amplification , isothermal process , mycobacterium , scale (ratio) , mycobacterium avium complex , microbiology and biotechnology , environmental chemistry , environmental science , chemistry , biology , bacteria , physics , thermodynamics , genetics , dna , quantum mechanics
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(®) 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.

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