Abstract

Data about the prevalence of the A2142C, A2142G, and A2143G mutations in 23S rRNA gene is still limited. The aim of this study was to determine the prevalence of these mutations in 23S rRNA gene of H. pylori vietnamese strains. One hundred and sixty-nine patients with H. pylori-positive chronic gastritis were examined. H. pylori was detected by rapid urease test and Polymerase chain reaction (PCR). Total DNA was extracted from gastric biopsy specimens. A2142C, A2142G, and A2143G mutations were detected by DNA sequencing and PCR-restriction fragment length polymorphism (PCR-RFLP). A2143G mutation was detected in 36.1% of samples, A2142G mutation in 3.6%, while A2142C mutation was not found in any case. The mixture of wild-type and mutation strains was found in 50% of specimens with A2142G, in 23% of specimens with A2143G mutation. There was no association of 23S rRNA gene point mutations with gender or age. However, an association between the heterogeneity of mutation and age was evidenced, with mean age of the group of pure A2143G higher than the group of wild-type/A2143G mixture, and rate of the wild-type/A2143G mixture higher in patients under 40 years of age. A2143G mutation was prominent, while A2142C mutation was not found in the 23S rRNA gene. PCR-RFLP has revealed a reliable assay allowing a rapid and cost-effective detection of clarithromycin-resistant strains. This is useful in countries as Vietnam with high prevalence of clarithromycin-resistance before choosing optimal therapy for H. pylori eradication.

Highlights

  • Data about the prevalence of the A2142C, A2142G, and A2143G mutations in 23S rRNA gene is still limited

  • The aim of our study was to determine the prevalence of clarithromycin-resistance among H. pylori isolates in Vietnamese patients with chronic gastritis via the detection of mutations at 2142 and 2143 positions of domain V of 23S rRNA gene by using DNA sequencing and Polymerase chain reaction (PCR)-restriction fragment length polymorphism (PCR-RFLP) on DNA extracted from gastric biopsy specimens

  • One hundred and eighty DNA samples were extracted from RUTpositive gastric biopsy specimens using Wizard Genomic DNA purification Kit (Promega, Madison, Wisconsin, USA), and H. pylori infection was confirmed by PCR using primers 5’AAGCTTTTAGGGGTGTTAGGGGTTT-3’ and 5’-AAGCTTACTTTCTAACACTAACGC-3’ targeting urease C gene [12]

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Summary

Introduction

Data about the prevalence of the A2142C, A2142G, and A2143G mutations in 23S rRNA gene is still limited. PCR-RFLP has revealed a reliable assay allowing a rapid and cost-effective detection of clarithromycin-resistant strains. This is useful in countries as Vietnam with high prevalence of clarithromycin-resistance before choosing optimal therapy for H. pylori eradication. In 1996, Versalovic observed the association of mutations at domain V of 23S rRNA gene with the status of clarithromycin resistance in strains of H. pylori [4]. These mutations are known as A2142G and A2143G, defined by Taylor et al in 1997 [5], which represent nowadays the molecular basis of the diagnosis of clarithromycin-resistant H. pylori through genotyping techniques. The mutations at 2142 and 2143 positions of 23S rRNA gene were responsible for more than 90% of clarithromycin resistance in developed countries [6], with the predominance of A2143G, A2142G and A2142C

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