The mutations that are responsible for fluoroquinolone resistance in the gyrA, gyrB, parC, and parE genes of Salmonella enterica serovar Typhi and serovar Paratyphi A were investigated. The sequences of the quinolone resistance-determining region of the gyrA gene in clinical isolates which showed decreased susceptibilities to fluoroquinolones had a single mutation at either the Ser-83 or the Asp-87 codon, and no mutations were found in the gyrB, parC, and parE genes.Fluoroquinolones have become the first-line drugs for the treatment of typhoid fever (3,12,18,23). However, some Salmonella enterica serovar Typhi strains that exhibit decreased susceptibilities to fluoroquinolones have been already reported (2,7,13,21). Furthermore, several clinical treatment failures after the administration of ciprofloxacin and other fluoroquinolones to patients with typhoid fever due to strains with decreased susceptibilities to fluoroquinolones have also been reported (17, 21). The emergence and spread of these organisms have been reported in developing countries. There is evidence that the incidence of strains that are resistant to nalidixic acid and that exhibit decreased susceptibilities to the most recent fluoroquinolones used for the treatment of typhoid fever is increasing. In most strains, the acquired fluoroquinolone resistance was attributed to mutations in the genes encoding DNA gyrase (GyrA, GyrB) (10, 24-26) or DNA topoisomerase IV (ParC, ParE) (8, 9). The purpose of this study was to investigate the association of quinolone resistance with mutations in the genes coding for gyrase and topoisomerase IV of S. enterica serovar Typhi and serovar Paratyphi A, which are especially clinically important serotypes of Salmonella spp.The bacterial strains used in this study were collected from regional public health offices in Japan between 1995 and 2001, and all isolates were obtained from a culture of either blood or stool from individual patients and identified by biochemical tests and serological tests on the basis of standard criteria. S. enterica serovar Typhi Ty2 and ATCC 19430 and S. enterica serovar Paratyphi A NCTC13, NCTC5702, and RIMD 1015 were used as reference and control strains (Tables 1 and 2). The MICs of several fluoroquinolones, including norfloxacin, levofloxacin, ofloxacin, sparfloxa cin, and ciprofloxacin, and nalidixic acid were determined by the Etest (AB Biodisk, Solna, Sweden), according to the instructions of the manufacturer. The quality control strains Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 were included in each test. The criterion for ciprofloxacin resistance was an MIC of Ն4 g/ml, according to the NCCLS breakpoint criteria for members of the family Enterobacteriaceae. The criterion for decreased susceptibility to ciprofloxacin that we used in this study was an MIC between Ն0.25 and Ͻ4 g/ml, and that for ciprofloxacin susceptibility was an MIC Ͻ0.25 g/ml (19,20). An attempt to increase the level of fluoroquinolone resistance was done by culturing the fluoroquinolone-susce...