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Background  Clarithromycin, amoxicillin, and a pump proton inhibitor are the most common drugs recommended as first-line triple therapy for  H.pylori  treatment, which results in eradication rates close to 80%, varying regionally, principally due to emergency cases and increases of clarithromycin resistant strains. Nucleotide substitutions at the  H. pylori  domain V of the 23S rRNA fraction are involved in the macrolide resistance and the A2142G and A2143G mutations are predominant in clinical isolates worldwide including in Brazil. As  H. pylori  culture is fastidious, we investigated the primary occurrence of  H. pylori  A2142G and A2143G rDNA 23S mutations using a molecular approach directly on gastric biopsies of dyspeptic patients consecutively attended at Hospital das Clinicas of Marilia, São Paulo, Brazil.    Methods  Biopsy specimens obtained from 1137 dyspeptic patients, were subjected to histopathology and  H. pylori  diagnosis by histology and PCR. PCR/RFLP assay was used to detect A2142G and A2143G point mutations at domain V of the  H. pylori  23S rDNA associated with clarithromycin resistance. Through the developed assay, a 768 bp PCR amplicon corresponding to1728 to 2495 bp of the 23S  H. pylori  rDNA is restricted with M boII  for A2142G mutation detection and with B saI  for A2143G mutation detection. Occurrence of 23S rDNA A2142G results in two DNA fragments (418 and 350 bp) and of 23S rDNA A2143G results in three DNA fragments (108, 310 and 350pb), due to a conserved B saI  restriction site.    Results  The PCR method used to diagnose  H. pylori  presented sensitivity, specificity and accuracy of 77,6%, 79,3% and 78,6%, respectively, compared to histology, the gold standard method for  H. pylori  diagnosis used in our routine. Prevalence of  H.pylori  with clarithromycin resistant genotypes was 2,46%, with predominance of A2143G 23S rDNA point mutation.    Conclusions  The PCR/RFLP assay was a rapid and accurate  H.pylori  diagnostic and clarithromycin resistance determination method useful for routine practice. As prevalence of primary resistance of  H.pylori  to clarithromycin due to A2142G and A2143G mutations remains low in Marilia, the standard clarithromycin containing triple therapy is still valid.

bmc gastroenterology

Low Helicobacter pylori primary resistance to clarithromycin in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil