Helicobacter pylori acquistion of metronidazole resistance by natural transformation in vitro.
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 8mg/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 ofH. 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.
Hua
Jie-Song
JS
Zheng
Peng-Yuan
PY
Fong
Teo Keng
TK
Khin
Mar Mar
MM
Bow
HO
H