Characterization of clinical isolates of Escherichia coli showing high levels of fluoroquinolone resistance.

During the years 1992 to 1994, an increase in fluoroquinolone-resistant Escherichia coli was observed at the Medical Center of the Technical University in Munich, Germany. Nineteen strains were collected and were thus available for further analysis. Pulsed-field gel electrophoresis showed clonal diversity in all but two strains. The majority of the patients from whom the strains were isolated had been previously treated with fluoroquinolones. Quinolone resistance was associated with mutations of the quinolone resistance-determining region of the gyrA gene in all cases. Direct sequencing of gyrA PCR amplification products revealed a mutation in codon 83 of the gyrA gene. In some instances the Ser-83-->Leu mutation was accompanied by an Asp-87-->Asn or Asp-87-->Gly mutation. Furthermore, the strains exhibited two different genotypes: in almost half of the fluoroquinolone-resistant E. coli strains as well as in the fluoroquinolone-susceptible E. coli reference strains ATCC 25922 and 35218, silent mutations were detected at bases 255, 273, 300, and 333. Although fluoroquinolones solved major problems in antimicrobial chemotherapy, in certain departments of our hospital the number of resistant E. coli isolates has become so high that susceptibility to fluoroquinolones can no longer be taken for granted.