Analysis by PCR and direct DNA sequencing of gyrA mutations associated with fluoroquinolone resistance in Enterococcus faecalis

A region of gyrA, the gene encoding subunit A of DNA gyrase, that is known to be associated with resistance was amplified and sequenced from 16 Enterococcus faecalis and Enterococcus faecium isolates. Six ciprofloxacin-resistant clinical isolates (MICs of ciprofloxacin, 32 to 64 micrograms/ml) and one multistep resistant laboratory mutant of E. faecalis (MIC of ciprofloxacin, 128 micrograms/ml) contained a change from serine to arginine or to isoleucine at codon 83 or a change from glutamic acid to lysine or to glycine at codon 87 (Escherichia coli GyrA coordinates); these changes have been associated with fluoroquinolone resistance in other species. No difference in the region studied was found in two ciprofloxacin-resistant E. faecium isolates (MICs, 32 micrograms/ml) or in four laboratory derived, spontaneous ciprofloxacin-resistant mutants of E. faecalis (MICs, 8 to 16 micrograms/ml), suggesting that other mechanisms may be responsible for fluoroquinolone resistance in some enterococci.