title: Development of a single multiplex amplification refractory mutation system PCR for the detection of rifampin-resistant Mycobacterium tuberculosis.
abstract: A rapid and simple method for the detection of drug-resistant Mycobacterium tuberculosis is critical for the efficient treatment and control of this pathogen in developing country. Here we developed a single multiplex amplification refractory mutation system (M-ARMS) PCR, in which chimeric-primer and temperature switch PCR (TSP) strategy were included. Using this method, we detected rifampin resistance-associated mutations at codons 511, 516, 526 and 531 in the rifampin resistance-determining region of rpoB gene. The performance of M-ARMS-PCR assay was evaluated with 135 cultured isolates of M. tuberculosis. The sensitivity and specificity were 94.2% and 100%, respectively, compared with direct DNA sequencing, and 86.67% and 89.71%, respectively, compared with culture-based phenotypic drug susceptibility testing. Therefore, this newly-developed M-ARMS-PCR method is useful and efficient with an intended application in provincial Centers for Disease Control and Prevention for rapid detection of rifampin resistance-associated mutations.
authors: Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changbai Rd 155, Beijing 102206, China; Department of Neurology, Xijing Hospital, The Fourth Military Medical University, Changle-Xi Road 167, Xi'an 710032, China.
authors: Xiaodan
authors: X
authors: Shi
authors: Chen
authors: C
authors: Zhang
authors: Ming
authors: M
authors: Shi
authors: Mengjie
authors: M
authors: Yang
authors: Yi
authors: Y
authors: Zhang
authors: Ji
authors: J
authors: Wang
authors: Hongwei
authors: H
authors: Shen
authors: Gang
authors: G
authors: Zhao
authors: Xuejun
authors: X
authors: Ma
mesh_terms: D004269:DNA, Bacterial / Q000187:drug effects; D024901:Drug Resistance, Multiple, Bacterial; D006801:Humans; D008826:Microbial Sensitivity Tests; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D012293:Rifampin / Q000627:therapeutic use*; D018088:Tuberculosis, Multidrug-Resistant / Q000235:genetics* / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; D012293:Rifampin
keywords: ARMS-PCR; DHHS; DR; DST; Department of Health and Human Services; Drug-resistant; EMB; INH; LJ; Lowenstein–Jensen media; M-ARMS-PCR; M. tuberculosis; MDR-TB; Mtb; Mycobacterium tuberculosis; RIF; RRDR; SDS; SM; amplification refractory mutation system PCR; drug resistance; drug susceptibility testing; ethambutol; isoniazid; multidrug-resistant tuberculosis; multiplex amplification refractory mutation system PCR; rifampicin; rifampicin resistance determining region; rpoB; sodium dodecyl sulfate; streptomycin