Multiplex allele-specific polymerase chain reaction for detection of isoniazid resistance in Mycobacterium tuberculosis

Int J Tuberc Lung Dis. 2011 Jun;15(6):799-803. doi: 10.5588/ijtld.10.0599.

Abstract

Setting: Pham Ngoc Thach Tuberculosis Reference Hospital, Ho Chi Minh City, Viet Nam.

Design: A multiplex allele-specific polymerase chain reaction (MAS-PCR) was developed to detect mutations at the two most common sites responsible for isoniazid (INH) resistance in Mycobacterium tuberculosis: katG315 and inhA-15. The MAS-PCR is able to detect rare mutations at katG315, in addition to katG S315T. Conventional phenotypic proportion drug susceptibility testing on Löwenstein-Jensen media was used as a gold standard to compare the sensitivity and specificity of the commercial MTBDRplus line-probe assay and the MAS-PCR in 100 INH-resistant and 50 INH-susceptible isolates collected consecutively at Pham Ngoc Thach Hospital reference laboratory.

Results: The sensitivity and specificity on culture isolates were 90% (n = 90/100, 95%CI 0.83-0.94) and 100% (n = 50/50, 95%CI 0.93-1.0), respectively, for the MAS-PCR and the MTBDRplus assay.

Conclusion: The MAS-PCR described here represents an alternative method for rapid screening for INH resistance in M. tuberculosis isolates.

MeSH terms

  • Alleles
  • Antitubercular Agents / pharmacology*
  • Bacterial Proteins / genetics
  • Catalase / genetics
  • DNA Primers
  • Drug Resistance, Bacterial / genetics*
  • Humans
  • Isoniazid / pharmacology*
  • Multiplex Polymerase Chain Reaction / economics
  • Multiplex Polymerase Chain Reaction / methods*
  • Mutation
  • Mycobacterium tuberculosis / drug effects
  • Mycobacterium tuberculosis / genetics*
  • Oxidoreductases / genetics
  • Sensitivity and Specificity
  • Sequence Analysis
  • Tuberculosis, Multidrug-Resistant / diagnosis
  • Tuberculosis, Multidrug-Resistant / drug therapy
  • Vietnam

Substances

  • Antitubercular Agents
  • Bacterial Proteins
  • DNA Primers
  • Oxidoreductases
  • Catalase
  • katG protein, Mycobacterium tuberculosis
  • InhA protein, Mycobacterium
  • Isoniazid