Helicobacter pylori acquisition of metronidazole resistance by natural transformation in vitro

Abstract

AIM: To study whether Helicobacter pylori is naturally transformable. METHODS: Transformation was performed in BHI broth supplemented with horse serum and yeast extract. Genomic DNA extracted from a metronidazole resistant H. pylori strain was added to H. pylori broth culture. The mixture was incubated at microaerophilic atmosphere. The DNA-treated cells were plated on blood agar containing 8 mg/ L metronidazole to select for transformants. Sterile distilled water was used as a negative DNA control. The DNA profiles of transformants were compared with that of their parent strains by randomly amplified polymorphic DNA (RAPD) fingerprinting. RESULTS: Transformation of H. pylori with DNA from a metronidazole resistant strain as a marker was demonstrated. Out of the 12 strains of H. pylori tested, 9 (75%) strains were found to be transformable. The transformation frequencies ranged from 3.4 × 10-6 to 2.4 × 10-4. By RAPD, DNA fingerprints of the transformants and their parent strains showed no change in DNA profiles though transformants were all resistant to metronidazole as compared with their metronidazole-sensitive parent strains. CONCLUSION: Helicobacter pylori is naturally transformable which might be one of the ways that H. pylori develops resistance to metronidazole.