The stethoscope, crucial in medical diagnosis, links doctors and patients. With the threat of hospital-acquired infections (HAIs), understanding stethoscope contamination and medical students' awareness is imperative. This study conducted from September to December 2022, the research involved 293 stethoscopes from three hospitals in the West Bank. Questionnaires probed students' practices, and Staphylococcus spp isolations were analyzed. Contamination rates, bacterial species, associations with disinfection practices and other hospital factors were explored. Stethoscope contamination rate was high in the three hospitals (range: 26.5–50.8 CFU/cm2). Staphylococcus spp contaminated 36.9% of stethoscopes. Methicillin Resistant S. aureus (MRSA) and Vancomycin Resistant S. aureus (VRSA) occurred in 16.6% and 1.8% respectively. 36% exhibited resistance to two or more antibiotics. The study revealed significant findings regarding stethoscope contamination. Those disinfecting stethoscopes between patients achieved a 29.4% acceptable rate, contrasting with a significantly higher 60.7% rate for non-disinfectors (P < 0.05). Reminders for disinfection in study units resulted in a 32.1% acceptable rate, whereas units without reminders had a significantly higher 67.9% rate (P < 0.001). Stethoscope usage also played a role, with a 34.4% acceptable rate for those examining only patients, compared to a significantly higher 65.8% rate for those examining both patients and peers (P < 0.05). Additionally, significant differences were observed in hospital, rotation, year-wise, disinfection frequency, and the presence of reminders. This study emphasizes the importance of stethoscope disinfection, reminders, and hand hygiene in reducing contamination. Educational interventions are critical.