A cohort study measured the occurrence and risk factors of nosocomial infections in the neonatal intensive care unit of Abha general hospital, Saudi Arabia. Of 401 neonates who stayed at least 48 hours in the unit, 77 developed infections, a period prevalence of 19.2% and an incidence of 13.7 infections per 1000 patient-days. The most frequent infections were: pneumonia (50.0%), primary bloodstream (40.9%) and skin and soft tissues (6.5%). In logistic regression analysis, mechanical ventilation (OR = 2.69, 95% CI: 1.39-5.19) and total parenteral nutrition (OR = 5.62, 95% CI: 2.78-11.35) were identified as significant risk factors. Neonates suffering from nosocomial infections had more than 3 times the risk of dying compared to neonates free of infection.