The gyrA gene mutations associated with quinolone resistance were determined in 21 epidemiologically unrelated clinical isolates of Acinetobacter baumannii. Our studies highlight the conserved sequences in the quinolone resistance-determining region of the gyrA gene from A. baumannii and other bacteria. All 15 isolates for which the MIC of ciprofloxacin is ≥4 μg/ml showed a change at Ser-83 to Leu. Six strains for which the MIC of ciprofloxacin is 1 μg/ml did not show any change at Ser-83, although u strain for which the MIC of ciprofloxacin is 1 μg/ml exhibited a change at Gly-81 to Val. Although it is possible that mutations in other locations of the gyrA gene, the gyrB gene, or in other genes may also contribute to the modulation of the MIC level, our results suggest that a gyrA mutation at Ser- 83 is associated with quinolone resistance in A. baumannii.