In this study, 154 Staphylococcus aureus isolates were detected from 1070 food samples (14.4%) collected from seven cities in Turkey. Antimicrobial susceptibility testing against 21 antibiotics was performed by agar disk diffusion method, and those isolates resistant to any antibiotic were further analyzed to determine minimum inhibitory concentration by E-test and polymerase chain reaction analysis of vanA and mecA genes. According to disk diffusion test results, a total of 139 strains were resistant to at least one tested antibiotic, with 39 (25.3%) strains being multidrug resistant (MDR) and the other 15 strains being susceptible to all antibiotics. Penicillin G, linezolid, erythromycin, and tetracycline took up 71.4%, 23.4%, 18.2%, and 15.6% of the tested strains, respectively. In addition, all of the strains were susceptible to vancomycin, oxacillin, cefoxitin, and imipenem. Only one strain (S158B) was resistant to both teicoplanin and cefazolin. On the other hand, the presence of vanA and mecA genes was not detected in the strains. Pulsed-field gel electrophoresis analysis was used to identify genetic-relatedness of the MDR strains. It is noteworthy that some strains from different sources showed 100% homology; however, some of MDR strains were found unrelated with 60% or less homology. The high diversity observed in pulsed-field gel electrophoresis results indicated the possible contamination of S. aureus from different sources and routes.