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

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 A926 G, whilst the five resistant isolates demonstrated double base pair substitutions of G927 T/A928 C and A926 G/A928 C and a single nucleotide substitution of A926 G.

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.