Simultaneous identification of rifampin-resistant Mycobacterium tuberculosis and nontuberculous mycobacteria by polymerase chain reaction-single strand conformation polymorphism and sequence analysis of the RNA polymerase gene (rpoB)

Bum-Joon Kim; Keun-Hwa Lee; Yeo-Jun Yun; Eun-Mi Park; Young-Gil Park; Gil-Han Bai; Chang-Yong Cha; Yoon-Hoh Kook

doi:10.1016/j.mimet.2004.03.009

Simultaneous identification of rifampin-resistant Mycobacterium tuberculosis and nontuberculous mycobacteria by polymerase chain reaction-single strand conformation polymorphism and sequence analysis of the RNA polymerase gene (rpoB)

J Microbiol Methods. 2004 Jul;58(1):111-8. doi: 10.1016/j.mimet.2004.03.009.

Authors

Bum-Joon Kim¹, Keun-Hwa Lee, Yeo-Jun Yun, Eun-Mi Park, Young-Gil Park, Gil-Han Bai, Chang-Yong Cha, Yoon-Hoh Kook

Affiliation

¹ Department of Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, and Clinical Research Institute, Seoul National University Hospital, Chongno-Gu, South Korea.

PMID: 15177909
DOI: 10.1016/j.mimet.2004.03.009

Abstract

Interspecies variations and mutations associated with rifampin resistance in rpoB of Mycobacterium allow for the simultaneous identification of rifampin-resistant Mycobacterium tuberculosis and nontuberculous mycobacteria by PCR-SSCP analysis and PCR- sequencing. One hundred and ten strains of rifampin-susceptible M. tuberculosis, 14 strains of rifampin-resistant M. tuberculosis, and four strains of the M. avium complex were easily identified by PCR-SSCP. Of another seven strains, which showed unique SSCP patterns, three were identified as rifampin-resistant M. tuberculosis and four as M. terrae complex by subsequent sequence analysis of their rpoB DNAs (306 bp). These results were concordant with those obtained by susceptibility testing, biochemical identification, and 16S rDNA sequencing.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Antitubercular Agents / pharmacology*
Base Sequence
DNA, Bacterial / chemistry
DNA, Bacterial / genetics
DNA-Directed RNA Polymerases / chemistry
DNA-Directed RNA Polymerases / genetics
Drug Resistance, Multiple, Bacterial
Molecular Sequence Data
Mycobacterium avium Complex / classification
Mycobacterium avium Complex / genetics
Mycobacterium tuberculosis / drug effects*
Mycobacterium tuberculosis / genetics
Mycobacterium tuberculosis / isolation & purification
Mycobacterium tuberculosis / metabolism
Phylogeny
Polymerase Chain Reaction
Polymorphism, Single-Stranded Conformational
Rifampin / pharmacology*
Sequence Alignment
Tuberculosis / microbiology

Substances

Antitubercular Agents
DNA, Bacterial
DNA-Directed RNA Polymerases
RNA polymerase beta subunit
Rifampin