Abstract: Staphylococcus aureus isolated from the skin surface of the athletes and their training environment in Malaysia were characterized for their antimicrobial susceptibility and Random Amplified Polymorphic DNA (RAPD). A total of 19 types of antibiotics used in this study and all the isolates were resistant to nalidixic acid and trimethoprim and showed Multiple Antibiotic Resistant (MAR) indexes, ranging from 0.11 to 0.68. No methicillin-resistant Staphylococcus aureus (MRSA) was found among the isolates tested in this study. The dendrogram obtained from the results of the RAPD-PCR discriminated the isolates into 4 clusters and 30 single isolates at the level of 100% similarity. Isolates from different individual and the environment were clustered together and colonized by similar strains.