title: Sub-speciation of Mycobacterium tuberculosis complex from tuberculosis patients in Japan.
abstract: Mycobacterium tuberculosis is the major causative agent of tuberculosis in humans. It is well known that Mycobacterium bovis and other species in the M. tuberculosis complex (MTC) can cause respiratory diseases as zoonosis. We analyzed the MTC isolates collected from tuberculosis patients from Japan in 2002 using a multiplex PCR system that detected cfp32, RD9 and RD12. A total of 970 MTC isolates that were representative of the tuberculosis cases throughout Japan, were examined using this method. As a result, 966 (99.6%) M. tuberculosis, two Mycobacterium africanum and two Mycobacterium canettii were identified using a multiplex PCR system, while no M. bovis was detected. Two isolates that lacked RD9 were initially considered to be M. canettii, but further analysis of the hsp65 sequence revealed them to be M. tuberculosis. Also two M. africanum were identified as M. tuberculosis using the -215 narG nucleotide polymorphism. Though PCR-linked methods have been used for a rapid differentiation of MTC and NTM, from our cases we suggest careful interpretation of RD based identification.
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan; Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan. Electronic address: ueyamam@fukujuji.org.
authors.forename: Masako
authors.initials: M
authors.lastname: Ueyama
authors.affiliation: Bacteriology Division, Department of Mycobacterium Reference and Research, Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Kinuyo
authors.initials: K
authors.lastname: Chikamatsu
authors.affiliation: Bacteriology Division, Department of Mycobacterium Reference and Research, Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Aono
authors.affiliation: Molecular Epidemiology Division, Department of Mycobacterium Reference and Research, Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Murase
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan; Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Kuse
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan; Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Kozo
authors.initials: K
authors.lastname: Morimoto
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan; Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Okumura
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yoshiyama
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Ogata
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan; Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Kozo
authors.initials: K
authors.lastname: Yoshimori
authors.affiliation: Department of Respiratory Medicine, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Kudoh
authors.affiliation: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Arata
authors.initials: A
authors.lastname: Azuma
authors.affiliation: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Gemma
authors.affiliation: Bacteriology Division, Department of Mycobacterium Reference and Research, Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Mitarai
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007564:Japan; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014376:Tuberculosis / Q000235:genetics* / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial
keywords: MTC differentiation; Multiplex-PCR; Mycobacterium africanum; Mycobacterium bovis; Mycobacterium tuberculosis complex