Staphylococcus aureus(S. aureus) is a ubiquitous pathogen responsible for several severe infections. This study aimed to investigate the adhesive properties and antibiotic resistance among clinicalS. aureusisolated from Hail Hospital Province, Kingdom of Saudi Arabia (KSA), using molecular approaches. This study was conducted according to the ethical committee at Hail’s guidelines on twenty-fourS. aureusisolates. A polymerase chain reaction (PCR) was performed to identify genes encoding the β-lactamase resistance (blaZ), methicillin resistance (mecA), fluoroquinolone resistance (norA), nitric oxide reductase (norB), fibronectin (fnbA andfnbB), clumping factor (clfA) and intracellular adhesion factors (icaA andicaD). This qualitative study tested adhesion based on exopolysaccharide production on Congo red agar (CRA) medium and biofilm formation on polystyrene byS. aureusstrains. Among 24 isolates, thecnaand blaz were the most prevalent (70.8%), followed bynorB (54.1%),clfA (50.0%),norA (41.6%),mecA andfnbB (37.5%) andfnbA (33.3%). The presence oficaA/icaD genes was demonstrated in almost all tested strains in comparison to the reference strain,S. aureusATCC 43300. The phenotypic study of adhesion showed that all tested strains had moderate biofilm-forming capacity on polystyrene and represented different morphotypes on a CRA medium. Five strains among the twenty-four harbored the four genes of resistance to antibiotics (mecA,norA,norB andblaz). Considering the genes of adhesion (cna,clfA,fnbA andfnbB), these genes were present in 25% of the tested isolates. Regarding the adhesive properties, the clinical isolates ofS. aureusformed biofilm on polystyrene, and only one strain (S17) produced exopolysaccharides on Congo red agar. All these results contribute to an understanding that the pathogenesis of clinicalS. aureusisolates is due to their antibiotic resistance and adhesion to medical material.