Spontaneous mutations that confer antibiotic resistance in Helicobacter pylori

In this study, we systematically examined in vitro frequencies and spectra of the spontaneous mutations in Helicobacter pylori that confer resistance to clarithromycin (Cla(r)), metronidazole (Mtz(r)), amoxicillin (Amx(r)), c iprofloxacin (Cip(r)), and rifampin (Rif(r)), The mutation rate of Rif(r) o r Cip(r) determined in a fluctuation assay is 1 x 10(-8) to 2 x 10(-8) per cell per division, In contrast, the mutation rates of Cla(r), Mtz(r), and A mx(r) are much lower (< 10(-9)). However, Mtz(r) mutants could be readily s elected in vitro by using the serial passage method, suggesting that the mu tagenic effect and selective effect of a sublethal dose of metronidazole co ntribute to the rapid development of Mtz(r). Analysis of spontaneous Rif(r) , Cla(r), and Cip(r) mutants confirmed previous results indicating that mut ations within the rpoB gene, the 23S rRNA gene, and the gyrA gene, respecti vely, are responsible; also, several new mutant alleles were identified. Mt z(r) mutants resulted most frequently, but not always, from mutations in th e rdxA gene. DNA fragments containing each mutant allele could readily tran sform susceptible H. pylori strains to resistance, confirming that each mut ant allele is responsible for the resistance phenotype.