title: Detection and identification of Mycobacterium tuberculosis in joint biopsy specimens by rpoB PCR cloning and sequencing.
abstract: Osteoarticular tuberculosis (OAT) is an extrapulmonary tuberculosis and accounts for 1 to 3% of all tuberculosis cases. We used an rpoB PCR-plasmid TA cloning-sequencing method to detect and identify tubercle bacilli in surgical specimens from patients suspected of having OAT. By comparing the similarities of the rpoB sequences determined with those in GenBank, Mycobacterium tuberculosis was detected in 23 of 43 samples. Three of the 23 positive samples had mutations at codon 531, which are commonly observed in rifampin-resistant M. tuberculosis strains. Our results suggest that the rpoB PCR-TA cloning-sequencing method developed, which detects M. tuberculosis and which simultaneously determines its rifampin susceptibility, can also be used efficiently for the diagnosis of OAT.
authors.affiliation: Department of Microbiology and Cancer Research Institute, Seoul National University College of Medicine, Seoul National Research Hospital, Korea.
authors.forename: Yeo-Jun
authors.initials: YJ
authors.lastname: Yun
authors.forename: Keun-Hwa
authors.initials: KH
authors.lastname: Lee
authors.forename: Lin
authors.initials: L
authors.lastname: Haihua
authors.forename: Yoon-Jong
authors.initials: YJ
authors.lastname: Ryu
authors.forename: Bum-Joon
authors.initials: BJ
authors.lastname: Kim
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Lee
authors.forename: Goo-Hyun
authors.initials: GH
authors.lastname: Baek
authors.forename: Hee-Joong
authors.initials: HJ
authors.lastname: Kim
authors.forename: Moon-Sang
authors.initials: MS
authors.lastname: Chung
authors.forename: Myung-Chul
authors.initials: MC
authors.lastname: Lee
authors.forename: Sang-Hoon
authors.initials: SH
authors.lastname: Lee
authors.forename: In-Ho
authors.initials: IH
authors.lastname: Choi
authors.forename: Tae-Jun
authors.initials: TJ
authors.lastname: Cho
authors.forename: Bong-Soon
authors.initials: BS
authors.lastname: Chang
authors.forename: Yoon-Hoh
authors.initials: YH
authors.lastname: Kook
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology; D001706:Biopsy; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D006801:Humans; D007596:Joints / Q000382:microbiology*; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012293:Rifampin / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D014394:Tuberculosis, Osteoarticular / Q000382:microbiology*
chemical_list: D000995:Antitubercular Agents; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D012293:Rifampin