Background: The aim of this study was to investigate the prevalence of plasmid-mediated quinolone resistance (PMQR) and mutations in quinolone resistance-determining regions (QRDRs) of Salmonella and their association with fluoroquinolone susceptibility in Korea. Methods: A total of 284 nonduplicated clinical isolates of Salmonella were collected from various clinical specimens at 12 tertiary-care hospitals in Korea. The qnrA, qnrB, and qnrS genes were detected by multiplex polymerase chain reaction (PCR). The qepA and aac(6′)-Ib-cr genes were amplified by PCR. The QRDRs of gyrA, gyrB, parC, and parE were amplified by PCR from the DNA of selected nalidixic acid-resistant and qnr-positive isolates. Results: We detected six qnr-positive Salmonella (four qnrS1 and two qnrB19) and one aac(6′)-Ib-cr-positive strain. A mutation in the QRDR of gyrA only (N=46) was the most common, followed by gyrA+parC (N=9), parC (N=7), gyrA+parE (N=3), parC+parE (N=3), gyrA+gyrB (N=2), and parE (N=1). There were seven novel mutations in the QRDR regions of gyrB, parC, and parE. Six of seven PMQR-positive isolates had high-level resistance to nalidixic acid, and all six strains had reduced susceptibility to ciprofloxacin. One qnrS1-positive isolate was resistant to ciprofloxacin, norfloxacin, and nalidixic acid. The resistant rates to nalidixic acid, ciprofloxacin, norfloxacin, and levofloxacin were 49.3%, 1.1%, 0.7%, and 0.4%, respectively. Conclusion: We report the first detection of PMQR in Salmonella isolates from Korea. It is essential to continue surveillance and to watch for the spread of PMQR in Salmonella for public health control.