title: Applicability of in-house loop-mediated isothermal amplification for rapid identification of Mycobacterium tuberculosis complex grown on solid media.
abstract: A simple, rapid, and low-cost identification method is required in tuberculosis high-burden countries. We report the applicability of in-house loop-mediated isothermal amplification (LAMP) targeting 16S ribosomal RNA for the rapid identification of Mycobacterium tuberculosis complex grown on Lowenstein-Jensen media. Eighty acid-fast staining-positive clinical isolates were selected and used to evaluate the LAMP assay in comparison with polymerase chain reaction and conventional culture-based tests. The LAMP assay identified 60 M. tuberculosis isolates from 80 clinical isolates using simple heat-extracted DNA directly from the colony suspension. The results were in complete agreement with those obtained using the other methods, and the utility of the direct LAMP assay from a colony was demonstrated. The LAMP assay appears to be a practical and low-cost method that can be used for the rapid identification of M. tuberculosis isolates and suitable for endemic low-resource settings.
authors.affiliation: National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.
authors.forename: Benjawan
authors.initials: B
authors.lastname: Phetsuksiri
authors.forename: Janisara
authors.initials: J
authors.lastname: Rudeeaneksin
authors.forename: Sopa
authors.initials: S
authors.lastname: Srisungngam
authors.forename: Supranee
authors.initials: S
authors.lastname: Bunchoo
authors.forename: Dhanida
authors.initials: D
authors.lastname: Roienthong
authors.forename: Tetsu
authors.initials: T
authors.lastname: Mukai
authors.forename: Chie
authors.initials: C
authors.lastname: Nakajima
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Hamada
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Suzuki
mesh_terms: D001431:Bacteriological Techniques / Q000191:economics / Q000379:methods*; D003365:Costs and Cost Analysis; D003470:Culture Media / Q000737:chemistry; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013997:Time Factors; D014376:Tuberculosis / Q000175:diagnosis*
chemical_list: D003470:Culture Media; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S