title: A simple, rapid and economic method for detecting multidrug-resistant tuberculosis.
abstract: OBJECTIVE
  To evaluate multiplex allele specific polymerase chain reaction as a rapid molecular tool for detecting multidrug-resistant tuberculosis.
  
  
  METHODS
  Based on drug susceptibility testing, 103 isolates were multidrug-resistant tuberculosis and 45 isolates were sensitive to isonicotinylhydrazine and rifampin. Primers were designed to target five mutations hotspots that confer resistance to the first-line drugs isoniazid and rifampin, and multiplex allele specific polymerase chain reaction was performed. Whole-genome sequencing confirmed drug resistance mutations identified by multiplex allele specific polymerase chain reaction.
  
  
  RESULTS
  DNA sequencing revealed that 68.9% of multidrug-resistant strains have point mutations at codon 315 of the katG gene, 19.8% within the mabA-inhA promoter, and 98.0% at three hotspots within rpoB. Multiplex allele specific polymerase chain reaction detected each of these five mutations, yielding 82.3% sensitivity and 100% specificity for isoniazid resistance, and 97.9% sensitivity and 100% specificity for rifampin resistance as compared to drug susceptibility testing.
  
  
  CONCLUSIONS
  The results show that multiplex allele specific polymerase chain reaction is an inexpensive and practical method for rapid detection of multidrug-resistant tuberculosis in developing countries.
authors.affiliation: Department of Tuberculosis, The First Affiliated Hospital of Xinxiang Medical University, Henan Province, China. Electronic address: xiawangcn@yeah.net.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.forename: Junhua
authors.initials: J
authors.lastname: Jiao
authors.forename: Weihua
authors.initials: W
authors.lastname: Xu
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Chai
authors.forename: Zhenyun
authors.initials: Z
authors.lastname: Li
authors.forename: Qingjiang
authors.initials: Q
authors.lastname: Wang
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D008826:Microbial Sensitivity Tests; D025202:Molecular Diagnostic Techniques; D060885:Multiplex Polymerase Chain Reaction / Q000191:economics; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D017354:Point Mutation; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D000995:Antitubercular Agents; D004269:DNA, Bacterial
keywords: Multiplex allele specific polymerase chain reaction (MAS-PCR); Mycobacterium tuberculosis
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