Microbial biofilms pose a significant danger to individuals needing medical devices (MDs). Extended durations of implantation present an elevated possibility of device related infections (DRIs). Staphylococcus bacteria are frequently detected in nosocomial infections related to medical devices due to their ability to adhere to the surface of biomaterials. This study aimed to assess the presence of Staphylococcus aureus in urinary, nasogastric, and peripheral venous catheters. Furthermore, this study examined the antibiotic resistance of Staphylococcus aureus isolated from MDs. Over six months, 62 samples were gathered from hospital devices in Khenchela, Algeria. S. aureus was identified using morphological and biochemical characteristics. Antibiotic resistance was assessed via disc diffusion following 30 isolates. Vancomycin was the most effective against S. aureus. The findings revealed a significant prevalence of multidrug-resistant bacteria in samples from biomaterial surfaces, which highlights the urgent need for advancements in hospital sterilization systems to curb the spread of the most resilient strains of S. aureus, thereby reducing the risk of device-related infections.