This study sought to analyze the antimicrobial resistant phenotypes and genotypes as well as the virulence content of isolates recovered from patients with diabetic foot infections (DFIs) in a Tunisian hospital. Eighty-three clinical samples of 64 patients were analyzed, and bacterial isolates were identified by MALDI-TOF. The antimicrobial resistance phenotypes were determined by the Kirby-Bauer disk diffusion susceptibility test. Resistance and virulence genes, profile, and SCC types were determined by PCR and sequencing. was detected in 14 of the 64 patients (21.9%), and 15 isolates were recovered. Six out of the fifteen isolates were methicillin-resistant (MRSA, -positive) (40%). The isolates harbored the following resistance genes (number of isolates): (12), (B) (2), (A) (1), (2), (M) (2), (K) (3), (L) (1), (6')(2″) (2), (4″) (1) and A (1). The and genes were detected in three isolates. Twelve different -types were identified and assigned to seven clonal complexes with the predominance of -type III. Furthermore, the SCC types III, IV and V were found among the MRSA isolates. Moreover, one MSSA CC398-t571--III isolate was found; it was susceptible to all antimicrobial agents and lacked and genes. This is the first report on the prevalence and molecular characterization of from DFIs and also the first detection of the MSSA-CC398-t571 clone in human infections in Tunisia. Our findings indicated a high prevalence in DFIs with genetic diversity among the MSSA and MRSA isolates.