title: Simultaneous detection of isoniazid, rifampin, and ethambutol resistance of Mycobacterium tuberculosis by a single multiplex allele-specific polymerase chain reaction (PCR) assay.
abstract: Prompt detection of drug resistance of Mycobacterium tuberculosis is essential for effective control of tuberculosis (TB). We developed a multiplex allele-specific polymerase chain reaction (MAS-PCR) that detects the most commonly observed isoniazid (INH), rifampin (RIF), and ethambutol resistance-associated mutations in a single assay. The usefulness of the newly developed method was evaluated with 174 clinical isolates of M. tuberculosis obtained from Turkey. Distinct PCR banding patterns were observed for different mutation profiles and the correlation between MAS-PCR results and DNA sequencing findings was 99.4%. With culture-based phenotypic drug susceptibility testing as a reference standard, the sensitivity and specificity of the newly developed MAS-PCR assay for drug resistance-related genetic mutation detection were determined to be 81.1% and 97.5% for INH, 93.0% and 98.9 % for RIF, and 54.5% and 68.0 % for ethambutol. MAS-PCR provides a rapid, potentially more cost-effective, method of detecting multidrug-resistant TB.
authors.affiliation: Department of Epidemiology, School of Public Health, University of Michigan, Ann Arbor, 48109-2029, USA. zhenhua@umich.edu
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Yang
authors.forename: Riza
authors.initials: R
authors.lastname: Durmaz
authors.forename: Dong
authors.initials: D
authors.lastname: Yang
authors.forename: Selami
authors.initials: S
authors.lastname: Gunal
authors.forename: Lixin
authors.initials: L
authors.lastname: Zhang
authors.forename: Betsy
authors.initials: B
authors.lastname: Foxman
authors.forename: Ahmet
authors.initials: A
authors.lastname: Sanic
authors.forename: Carl F
authors.initials: CF
authors.lastname: Marrs
mesh_terms: D000483:Alleles; D000995:Antitubercular Agents / Q000494:pharmacology*; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004977:Ethambutol / Q000494:pharmacology*; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012293:Rifampin / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D000995:Antitubercular Agents; D003470:Culture Media; D004269:DNA, Bacterial; D004977:Ethambutol; D007538:Isoniazid; D012293:Rifampin