Bacterial infections are an important cause of neonatal mortality and morbidity. The major pathogens for neonatal sepsis in the neonatal intensive care unit (NICU) vary with geographical area and time. It is therefore important to frequently audit neonatal sepsis in individual NICU, to aid in provision of adequate and appropriate preventive and therapeutic measures. We retrospectively reviewed the medical records of all infants who had positive blood cultures during a 2-year period in the NICU at a university hospital in Riyadh, Saudi Arabia. Overall the incidence of proven-bacterial-sepsis (PBS) was 10.2% of NICU admissions. The incidence of PBS in low-birth-weight (LBW), very low-birth-weight (VLBW), and extremely low-birth-weight (ELBW) infants were 19%, 41%, and 49% respectively. Multiple episodes of bacterial sepsis occurred in 21% of all infants infected. Coagulase negative Staphylococcus (CONS) (50%) was the most common infecting organism causing late onset sepsis (LOS) and Escherichia coli (29%) the most common causing early onset sepsis (EOS). Gram negative bacteria (GNB) were the infecting organisms in 50% of the EOS episodes and 29% of LOS episodes. Only 11% (14) of the PBS were EOS. Only 10 (10.4%) infants had bacterial meningitis. The overall PBS related mortality was 9%, representing 22% of all neonatal deaths.