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Objectives: The aim of our study was to evaluate the frequency of osteoarticular infections with Panton-Valentine leukocidin-positive (PVL) Staphylococcus aureus (PVL-SA) among patients admitted to the orthopedic ward at the Sahloul University Hospital (Sousse, Tunisia) and to study the characteristics of these strains and patients., Materials and Methods: We conducted a retrospective descriptive study over a 5-year period. Bacterial identification, antibiotic susceptibility, and molecular study (PCR to detect of the luk-PV gene that encodes PVL) were performed for 44 S. aureus isolates., Results: Panton-Valentine toxin was found in 41% of S. aureus cases, mainly males, and 39% of the PVL(+) cases were methicillin-sensitive (MSSA). These strains constitute a reservoir of PVL genes that can lead to the emergence and spread of PVL-SA clones resistant to methicillin (MRSA). In our series, PVL-MRSA accounted for 9% of all S. aureus isolates. Their profile and antibiotic resistance is that of clone ST80, frequently isolated in Europe and also reported in Algeria and Tunisia., Conclusion: It is desirable to test for PVL routinely in the laboratory to implement appropriate treatment and to monitor the epidemiology of these PVL-SA strains actively. Further measures should be undertaken to prevent and fight infections by these strains.