title: Direct detection of Mycobacterium avium in environmental water and scale samples by loop-mediated isothermal amplification.
abstract: 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.
authors.affiliation: Toneyama Institute for Tuberculosis Research, Osaka City University Medical School, 5-1-1 Toneyama, Toyonaka, Osaka 560-8552, Japan E-mail: nishiuchi@med.osaka-cu.ac.jp.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Nishiuchi
authors.affiliation: Department of Infectious Diseases, The Osaka Prefectural Institute of Public Health, 1-3-69 Nakamichi, Higashinari-ku, Osaka 537-0025, Japan.
authors.forename: Aki
authors.initials: A
authors.lastname: Tamaru
authors.affiliation: Research Center for Zoonosis Control, Hokkaido University, North 20, West 10 Kita-ku, Sapporo, Hokkaido 001-0020, Japan.
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: National Hospital Organization Toneyama National Hospital, 5-1-1 Toneyama, Toyonaka, Osaka 560-8552, Japan.
authors.forename: Seigo
authors.initials: S
authors.lastname: Kitada
authors.affiliation: National Hospital Organization Toneyama National Hospital, 5-1-1 Toneyama, Toyonaka, Osaka 560-8552, Japan.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Maekura
authors.affiliation: National Hospital Organization Toneyama National Hospital, 5-1-1 Toneyama, Toyonaka, Osaka 560-8552, Japan; Department of Bacteriology, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-8585, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Tateishi
authors.affiliation: Department of Bacteriology, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-8585, Japan.
authors.forename: Mamiko
authors.initials: M
authors.lastname: Niki
authors.affiliation: Department of Virology, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-8585, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Ogura
authors.affiliation: Department of Bacteriology, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-8585, Japan.
authors.forename: Sohkichi
authors.initials: S
authors.lastname: Matsumoto
mesh_terms: D004784:Environmental Monitoring / Q000379:methods*; D006801:Humans; D007564:Japan; D015269:Mycobacterium avium Complex / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D015270:Mycobacterium avium-intracellulare Infection / Q000382:microbiology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010362:Patients' Rooms; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity; D014871:Water Microbiology
chemical_list: D012336:RNA, Ribosomal, 16S