Molecular analysis of low-level tetracycline resistance in clinical isolates of Helicobacter pylori among dyspeptic patients in South West Nigeria

J Glob Antimicrob Resist. 2018 Jun:13:143-145. doi: 10.1016/j.jgar.2018.01.003. Epub 2018 Jan 31.

Abstract

Objectives: The aim of this study was to determine the occurrence of 16S rRNA mutations associated with low-level tetracycline resistance in Helicobacter pylori isolates from adult dyspeptic patients in South West Nigeria.

Methods: Susceptibility testing to tetracycline of 50 H. pylori isolates was performed by Etest. The 535-bp conserved region of the H. pylori tetracycline-binding site of 16S rRNA was amplified by PCR, followed by sequencing and multiple sequence alignment for all 50 clinical isolates.

Results: Of the 50 clinical isolates examined, DNA sequence analysis revealed nucleotide substitutions in 7 isolates at positions 926-928. Of the seven isolates, two demonstrated reduced susceptibility to tetracycline with Etest minimum inhibitory concentrations (MICs) of 0.75-1.0mg/L, whilst the other five isolates were resistant with MICs of 1.5-24mg/L (resistance breakpoint >1mg/L). The two isolates with reduced susceptibility had single nucleotide substitution of A926G, whilst the five resistant isolates demonstrated double base pair substitutions of G927T/A928C and A926G/A928C and a single nucleotide substitution of A926G.

Conclusions: This study shows that low-level tetracycline resistance amongst H. pylori-positive dyspeptic patients is associated with reduced susceptibility and resistance to tetracycline. This is the result of 1-bp and 2-bp differences in positions 926 and 926-928, respectively, in the 16S rRNA of H. pylori.

Keywords: Helicobacter pylori; Low-level resistance; Nigeria; Susceptibility; Tetracycline.

MeSH terms

  • Anti-Bacterial Agents
  • Biopsy
  • DNA, Bacterial / genetics
  • Dyspepsia / epidemiology
  • Dyspepsia / microbiology*
  • Helicobacter Infections / drug therapy
  • Helicobacter Infections / epidemiology
  • Helicobacter Infections / microbiology
  • Helicobacter pylori / drug effects*
  • Helicobacter pylori / isolation & purification
  • Humans
  • Microbial Sensitivity Tests
  • Mutation
  • Nigeria / epidemiology
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA
  • Stomach / microbiology
  • Stomach / pathology
  • Tetracycline / pharmacology*
  • Tetracycline / therapeutic use
  • Tetracycline Resistance / genetics*

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • RNA, Ribosomal, 16S
  • Tetracycline