Objective To investigate the antimicrobial susceptibility of 97 clinical Staphylococcus aureus (S. aureus) strains against 14 antimicrobials and corresponding resistance mechanisms. Methods The antimicrobial susceptibility of the isolates was determined using a disk diffusion method and antimicrobial resistance genes were screened by polymerase chain reaction. Mutations responsible for ciprofloxacin and rifampicin resistance were investigated by polymerase chain reaction and DNA sequencing. Results All isolates were found to be susceptible to vancomycin. Various rates of resistance to penicillin (83.5%), ampicillin (77.3%), erythromycin (63.9%), tetracycline (16.5%), amoxicillin/clavulanic acid (16.5%), ciprofloxacin (15.5%), trimethoprim/sulfamethoxazole (15.5%), oxacillin (13.4%), fusidic acid (12.4%), rifampin (6.2%), clindamycin (6.2%), gentamicin (6.2%) and mupirocin (5.2%) were determined. In addition, different combinations of resistance genes were identified among resistant isolates. Ciprofloxacin resistant isolates had mutations in codon 84 (Ser84Leu) and 106 (Gly106Asp) in the gyrA gene. Mutations in grlA were mostly related to Ser80Phe substitution. Leu466Ser mutation in the rpoB gene was detected in all rifampin resistant isolates. All methicillin resistant S. aureus isolates were SCCmec type V. Conclusions In conclusion, it was determined that the isolates were resistant to different classes of antimicrobials at varying rates and resistance was mediated by different genetic mechanisms. Therefore, continuous monitoring of resistance in S. aureus strains is necessary to control their resistance for clinically important antimicrobials.