A collection of 68 isolates of Salmonella enterica serovar Kentucky collected during the period 2003-2004 from humans in two geographical regions in the Slovak Republic was studied. The original isolate of this serovar was associated with travel to Egypt, and the emergence of other isolates was due to the nosocomial spread of this strain in two hospitals. Antimicrobial susceptibility testing, class I integrons content, pulsed-field gel electrophoresis (PFGE) analysis and plasmid DNA profiles were performed on all isolates. A high proportion (89.7%) of the isolates was multidrug-resistant, while 67 strains expressed resistance against ciprofloxacin. By sequence analysis of randomly selected strains, the point mutations in quinolone resistance-determining region of the DNA gyrase were found. The S. Kentucky isolates investigated were determined to be clonally related by PFGE as well as plasmid DNA analysis.