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Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis
Published on: May 16, 2025
Molecular Patterns of Resistance Among Helicobacter pylori Strains in South-Western Poland
Aldona Bińkowska1, Monika Maria Biernat2, Łukasz Łaczmański3
12nd Military Field Hospital of the Polish Armed Forces, Wrocław, Poland.
Abstract:
Treatment failure of Helicobacter pylori infection is caused mainly by progressive antibiotic resistance among H. pylori strains. In Poland, the prevalence of H. pylori strains resistant to metronidazole is higher than in other developed countries, reaching almost 50%, and resistance to clarithromycin is as high as 30% and is still increasing, contributing to the failure of first-line therapy in approximately 70% of patients. Moreover, the introduction of levofloxacin to eradication therapy of H. pylori infection quickly led to the emergence of resistant strains. Therefore, a necessary approach in microbiological diagnostics of H. pylori infection should be determination of susceptibility of H. pylori strains before the eradication treatment. Aim: In this study was to evaluate the molecular mechanisms of resistance among 170 H. pylori strains to clarithromycin, involving mutations in the 23S rRNA gene (A2143G, A2142G, A2143G) and to levofloxacin, involving mutations of gyrA and gyrB. Analysis was performed by using polymerase chain reaction and classical sequencing of DNA fragments. Results: Among examined strains, 26% were fully sensitive and 74% were resistant to at least one of the tested antibiotics. The overall resistance rate to metronidazole was as high as 56%, whereas to clarithromycin 46%, respectively. Resistance to LEV occurred among 6% of strains. All tested strains were susceptible to AMC and TET. The A2143G point mutation was found in 72% of clarithromycin-resistant strains. The most common mutation, present in 40% of H. pylori strains resistant to levofloxacin, was a change at position 91 of gyrA. Conclusion: The increasing number of point mutations in the 23S rRNA gene leads to an increase in the rates of antimicrobial resistance. Presence of the GCG allele at position 122 of the gyrA gene may cause an eightfold increase in risk of development of resistance to levofloxacin.
Insights
Antibiotic resistance in Helicobacter pylori strains, particularly to clarithromycin and metronidazole, is increasing in Poland, leading to treatment failures. Molecular analysis revealed specific gene mutations driving this resistance, underscoring the need for susceptibility testing before treatment.
Area of Science:
- Medical microbiology
- Molecular biology
- Infectious diseases
Background:
- Treatment failure in Helicobacter pylori infections is primarily due to rising antibiotic resistance.
- Poland exhibits high rates of H. pylori resistance to metronidazole (nearly 50%) and clarithromycin (30%), contributing to ~70% first-line therapy failure.
- Emergence of levofloxacin resistance in H. pylori strains necessitates pre-treatment susceptibility determination.
Purpose of the Study:
- To investigate molecular mechanisms of clarithromycin and levofloxacin resistance in 170 H. pylori strains.
- To identify specific mutations in the 23S rRNA gene (for clarithromycin) and gyrA/gyrB genes (for levofloxacin).
Main Methods:
- Polymerase chain reaction (PCR) and classical DNA sequencing were employed.
- Analysis focused on mutations A2143G, A2142G in 23S rRNA and mutations in gyrA/gyrB genes.
Main Results:
- 74% of H. pylori strains showed resistance to at least one antibiotic; overall resistance rates were 56% for metronidazole and 46% for clarithromycin.
- Levofloxacin resistance was observed in 6% of strains. All strains were susceptible to amoxicillin/clavulanic acid (AMC) and tetracycline (TET).
- The A2143G mutation in 23S rRNA was present in 72% of clarithromycin-resistant strains; a gyrA mutation at position 91 was most common (40%) in levofloxacin-resistant strains.
Conclusions:
- Increasing point mutations in the 23S rRNA gene correlate with rising antimicrobial resistance rates.
- The GCG allele at position 122 of the gyrA gene may significantly increase the risk (eightfold) of developing levofloxacin resistance.
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Resistivity
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