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A simple, rapid and economic method for detecting multidrug-resistant tuberculosis.

AbstractOBJECTIVE:
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.
AuthorsXia Wang, Junhua Jiao, Weihua Xu, Xiaoyan Chai, Zhenyun Li, Qingjiang Wang
JournalThe Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases (Braz J Infect Dis) 2013 Nov-Dec Vol. 17 Issue 6 Pg. 667-71 ISSN: 1678-4391 [Electronic] Brazil
PMID24029439 (Publication Type: Journal Article)
CopyrightCopyright © 2013 Elsevier Editora Ltda. All rights reserved.
Chemical References
  • Antitubercular Agents
  • DNA, Bacterial
Topics
  • Antitubercular Agents (pharmacology)
  • DNA, Bacterial (analysis)
  • Humans
  • Microbial Sensitivity Tests
  • Molecular Diagnostic Techniques
  • Multiplex Polymerase Chain Reaction (economics)
  • Mycobacterium tuberculosis (drug effects, genetics)
  • Point Mutation
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Tuberculosis, Multidrug-Resistant (diagnosis, microbiology)

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