Detection and identification of Mycobacterium tuberculosis in joint biopsy specimens by rpoB PCR cloning and sequencing.
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.
Yun
Yeo-Jun
YJ
Department of Microbiology and Cancer Research Institute, Seoul National University College of Medicine, Seoul National Research Hospital, Korea.
Lee
Keun-Hwa
KH
Haihua
Lin
L
Ryu
Yoon-Jong
YJ
Kim
Bum-Joon
BJ
Lee
Young-Ho
YH
Baek
Goo-Hyun
GH
Kim
Hee-Joong
HJ
Chung
Moon-Sang
MS
Lee
Myung-Chul
MC
Lee
Sang-Hoon
SH
Choi
In-Ho
IH
Cho
Tae-Jun
TJ
Chang
Bong-Soon
BS
Kook
Yoon-Hoh
YH
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*
D000995:Antitubercular Agents; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D012293:Rifampin