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Mutations of rpoB, katG, inhA and ahp genes in rifampicin and isoniazid-resistant Mycobacterium tuberculosis in Kyrgyz Republic

BMC Microbiol. 2018 Mar 22;18(1):22. doi: 10.1186/s12866-018-1168-x.

Abstract

Background: The aim of this study was to identify mutations of rpoB, katG, inhA and ahp-genes associated Mycobacterium tuberculosis resistance to rifampicin (RIF) and isoniazid (INH) in Kyrgyz Republic. We studied 633 smear samples from the primary pulmonary tuberculosis (TB) patients. We verified Mycobacterium tuberculosis susceptibility to RIF and INH using culture method of absolute concentrations, and commercially available test named "TB-BIOCHIP" (Biochip-IMB, Moscow, Russian Federation).

Results: For RIF-resistance, TB-BIOCHIP's sensitivity and specificity were 88% and 97%, 84% and 95% for INH-resistance, and 90% and 97% for multi-drug resistance (MDR). In RIF-resistant strains, TB-BIOCHIP showed mutations in codons 531 (64.8%), 526 (17.3%), 516 (8.1%), 511 (5.4%), 533 (3.2%), 522 (0.6%) and 513 (0.6%) of rpoB gene. The most prevalent was Ser531 > Leu mutation (63.7%). 91.2% of mutations entailing resistance to INH were in katG gene, 7% in inhA gene, and 1.8% in ahpC gene. Ser315→Thr (88.6%) was the most prevalent mutation leading to resistance to INH.

Conclusions: In Kyrgyz Republic, the most prevalent mutation in RIF-resistant strains was Ser531 → Leu in rpoB gene, as opposed to Ser315 → Thr in katG gene in INH-resistant Mycobacterium tuberculosis. In Kyrgyz Republic, the major reservoir of MDR Mycobacterium tuberculosis were strains with combined mutations Ser531 → Leu in rpoB gene and Ser315 → Thr in katG gene. TB-BIOCHIP has shown moderate sensitivity with the advantage of obtaining results in only two days.

Keywords: Ahp; Kyrgyz Republic; Mycobacterium tuberculosis; TB-BIOCHIP; inhA; katG; rpoB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Antitubercular Agents / pharmacology*
  • Bacterial Proteins / genetics*
  • Catalase / genetics*
  • DNA, Bacterial / genetics
  • DNA-Directed RNA Polymerases / genetics*
  • Drug Resistance, Multiple, Bacterial / drug effects
  • Drug Resistance, Multiple, Bacterial / genetics
  • Female
  • Humans
  • Isoniazid / pharmacology
  • Kyrgyzstan / epidemiology
  • Male
  • Microbial Sensitivity Tests
  • Middle Aged
  • Molecular Epidemiology
  • Mutation Rate
  • Mutation*
  • Mycobacterium tuberculosis / drug effects
  • Mycobacterium tuberculosis / genetics*
  • Oxidoreductases / genetics*
  • Peroxidases / genetics
  • Phenotype
  • Rifampin / pharmacology
  • Tuberculosis, Multidrug-Resistant / genetics*
  • Tuberculosis, Multidrug-Resistant / microbiology
  • Young Adult

Substances

  • Antitubercular Agents
  • Bacterial Proteins
  • DNA, Bacterial
  • rpoB protein, Mycobacterium tuberculosis
  • Oxidoreductases
  • Peroxidases
  • Catalase
  • katG protein, Mycobacterium tuberculosis
  • InhA protein, Mycobacterium
  • DNA-Directed RNA Polymerases
  • Isoniazid
  • Rifampin