Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR

Hiroaki Inoue; Reiko Fujimura; Kunio Agata; Hiroyuki Ohta

doi:10.1264/jsme2.ME14115

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Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR

Hiroaki Inoue, Reiko Fujimura, Kunio Agata, Hiroyuki Ohta

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Keywords: clone library, Ethidium monoazide (EMA), Legionella, quantitative polymerase chain reaction (qPCR), Viable but nonculturable (VBNC)

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Supplementary material

2015 Volume 30 Issue 1 Pages 108-112

DOI https://doi.org/10.1264/jsme2.ME14115

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Abstract

Viable Legionella spp. in environmental water samples were characterized phylogenetically by a clone library analysis combining the use of ethidium monoazide and quantitative PCR. To examine the diversity of Legionella spp., six cooling tower water samples and three bath water samples were collected and analyzed. A total of 617 clones were analyzed for their 16S rRNA gene sequences and classified into 99 operational taxonomic units (OTUs). The majority of OTUs were not clustered with currently described Legionella spp., suggesting the wide diversity of not-yet-cultured Legionella groups harbored in cooling tower water environments.

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