title: Distribution of rpoB mutations among multidrug-resistant Mycobacterium tuberculosis (MDRTB) strains from Thailand and development of a rapid method for mutation detection.
abstract: Since rifampicin resistance is a surrogate marker for multidrug-resistant Mycobacterium tuberculosis (MDRTB), the present study aimed to investigate rpoB mutations conferring rifampicin resistance in M. tuberculosis strains from Thailand, and to develop a rapid, inexpensive and simple PCR-based method for rapid detection of MDRTB. Overall, 267 M. tuberculosis isolates, including 143 MDRTB isolates, were investigated. Isolates of the Beijing strain predominated among the MDRTB isolates (79.1%), but accounted for only 45.5% of the susceptible isolates. Mutations in the rpoB gene were found most commonly at codons 531, 526 and 516 (58%, 25.2% and 9.1%, respectively). A multiplex allele-specific PCR was developed and tested with 216 clinical isolates. In comparison with the proportion method, the method showed 94.2% sensitivity and 100% specificity, and had a 100% positive predictive value and a 95% negative predictive value, which suggested that this method could be useful for screening for MDRTB, particularly in resource-limited countries.
authors.affiliation: National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Thailand Science Park, Pathumthani, Thailand.
authors.forename: T
authors.initials: T
authors.lastname: Prammananan
authors.forename: W
authors.initials: W
authors.lastname: Cheunoy
authors.forename: D
authors.initials: D
authors.lastname: Taechamahapun
authors.forename: J
authors.initials: J
authors.lastname: Yorsangsukkamol
authors.forename: S
authors.initials: S
authors.lastname: Phunpruch
authors.forename: P
authors.initials: P
authors.lastname: Phdarat
authors.forename: M
authors.initials: M
authors.lastname: Leechawengwong
authors.forename: A
authors.initials: A
authors.lastname: Chaiprasert
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D012293:Rifampin / Q000494:pharmacology; D013785:Thailand
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C505805:rpoB protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin