Analysis of antimicrobial resistance patterns and genetic mutations in Helicobacter pylori from West Bengal, India depicting escalating clarithromycin and high levofloxacin resistance
Abstract
BACKGROUND: The growing resistance of Helicobacter pylori (H. pylori) to antibiotics poses a significant global health challenge, particularly in developing regions where infection rates are high, and routine antibiotic susceptibility surveillance is limited. This study aimed to evaluate the antimicrobial resistance patterns and associated genetic mutations in H. pylori strains isolated from patients in West Bengal (WB), India. METHODS: A total of 88 H. pylori strains were isolated from gastric biopsy samples collected between 2018 and 2020 from patients diagnosed with gastritis, duodenal ulcer, or gastric cancer. Antimicrobial susceptibility was determined using the agar dilution method against six antibiotics: metronidazole, tetracycline, clarithromycin, furazolidone, levofloxacin, and amoxicillin. Resistance-associated mutations in gyrA (levofloxacin), frxA and rdxA (metronidazole), and pbp1A (amoxicillin) were characterized via Sanger sequencing. Mutations in the 23SrRNA gene, implicated in clarithromycin resistance, were identified through allele-specific PCR, validated by Sanger sequencing and transformation assays. Additionally, BsaI and BbsI restriction enzyme digestion was used to confirm specific point mutations in 23SrRNA gene of clarithromycin-resistant isolates. RESULTS: Of 210 biopsies analyzed, 79 (37.6%) were H. pylori-positive, yielding 88 distinct strains-some patients harbored multiple genetically diverse isolates, identified by differences in cagA/vacA…
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