Background: Not only is it crucial to rapidly detect Staphylococcus epidermidis (S. epidermidis) isolates from a broad range of bacteria, but recognizing resistance agents can greatly improve current diagnostic and therapeutic strategies. Methods: The current cross-sectional study investigated 120 clinical isolates from a nosocomial S. epidermidis infection. The isolates were identified using common biochemical tests, and specific S. epidermidis surface protein C (SesC) primers were used to confirm the presence of S. epidermidis. PCR and special primers were used to detect the β,-lactamase gene (blaZ). Methicillin resistance was measured using the agar screening method and antibiotic susceptibility was measured by disk diffusion. Results: 100 samples were characterized as S. epidermidis using a phenotypic and genotypic methods. From the 100 specimens examined, 80% contained blaZ. According to agar screening, 60% of isolates were methicillin-resistant. S. epidermidis isolates demonstrated the highest resistance to penicillin (93%) and the highest sensitivity to cefazolin (39%). Conclusions: The increased resistance to β,-lactam antibiotics in S. epidermidis isolates is alarming, and certain precautions should be taken by healthcare systems to continuously monitor the antimicrobial pattern of S. epidermidis, so that an appropriate drug treatment can be established.