Background: Staphylococcus aureus strains that are Panton–Valentine leukocidin (PVL) positive cause severe skin and soft tissue infections as well as necrotizing pneumonia. The presence of PVL gene is a marker for methicillin-resistant S. aureus; therefore, survey on prevalence and phylogenetic distribution of PVL is of great importance for public health. The aim of this research was molecular epidemiology survey of S. aureus PVL positive, isolated from two tertiary hospitals of Sanandaj. Materials and Methods: A total of 264 staphylococci isolates were collected from clinical specimens, hospital personnel and hospital environment of two tertiary hospitals of Sanandaj, in 2012 (Toohid and Besat). Bacterial cultures and biochemical tests were performed for S. aureus detection. Then, polymerase chain reaction (PCR) and repetitive sequence-based PCR (rep-PCR) were used for the determination of prevalence and molecular epidemiology of S. aureus PVL, respectively. Data were analyzed using the Fisher's exact test (P < 0.05). Results: From 264 staphylococci isolates, 88 (33.33%) were detected as S. aureus. Furthermore, 20 out of 88 (22.72%) strains of S. aureus were PVL positive according to PCR results. Rep-PCR showed six main clusters of S. aureus samples. PVL had similar clonality between different samples. No significant relationship was observed between PVL positive S. aureus and rep-PCR patterns (P = 0.98). Conclusion: These results showed that a clone of S. aureus PVL positive has spread between the community and hospital settings. Therefore, appropriate measures are required to prevent the spread of staphylococci and other bacteria in hospitals.