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Information on the prevalence of S. aureus and the current antimicrobial resistance profile is necessary in selecting the appropriate treatment of S. aureus infections in any part of the world. This study examined the frequency and antibiotic resistance list of S. aureus isolates obtained from clinical specimens at one hospital in Korea. A total of 1,746 gram positive cocci collected were identified as S. aureus. S. aureus isolates were obtained from different samples including sputum (N=565; 32.4%), endotracheal aspirate (358; 20.5%), wounds (329; 18.8%), blood (137; 7.8%), urine (67; 3.8%), and pus (59; 3.4%). All 1,545 S. aureus (100%) strains screened from sputum (565; 36.6%), endotracheal aspirate (388; 25.1%), wounds (329; 21.3%), blood (137; 8.9%), urine (67; 4.3%), and pus (59; 3.8%) were sensitive to glycopeptide (vancomycin, teicoplanin), oxazolidinone (linezolid) and stretogramin (quinupristin/dalfopristin). The prevalence of resistant S. aureus was significantly (P＜0.01) lower in urine, blood, pus, wounds, and sputum than in endotracheal aspirates. As a result, there was a significant difference in the antibiotic resistance of S. aureus according to the clinical specimens.