Staphylococcus aureusis both a human commensal and a pathogen, that causes serious nosocomial and community-acquired infections. Despite nostrils being considered its preferred host habitat, the oral cavity has been demonstrated to be an ideal starting point for auto-infection and transmission. The antibiotic resistance assessment ofS. aureusis a priority and is often reported in clinical settings. This study aimed to explore the prevalence and antimicrobial susceptibility ofS. aureusin the oral and nasal cavities of healthy individuals. The participants (n= 101) were subjected to a demographic and clinical background survey, a caries evaluation, and to oral and nasal swabbing. Swabs were cultured in differential/selective media andS. aureusisolates were identified (MALDI-TOF MS) and tested for antibiotic susceptibility (EUCAST/CLSI). SimilarS. aureusprevalence was found exclusively on nasal (13.9%) or oral (12.0%) habitats, whereas 9.9% of the population were simultaneous nasal and oral carriers. In oro-nasal cavities, similar antibiotic resistance rates (83.3–81.5%), including MDR (20.8–29.6%), were observed. Notably, 60% (6/10) of the simultaneous nasal and oral carriers exhibited different antibiotic resistance profiles between cavities. This study demonstrates the relevance of the oral cavity as an independent colonization site forS. aureusand as a potential source of antimicrobial resistance, a role which has been widely neglected so far.