Background: Nosocomial infections (NIs) have become a matter of major concern in neonatal intensive care units (NICUs). Nosocomial infections are the result of the interaction of several risk factors. Objective: to identify risk factors for nosocomial infections among neonates admitted to neonatal intensive care unit in Pediatric Assiut University Hospital (AUH). Knowledge of modifiable risk factors could be used to guide the design of interventions to prevent the problem. Methods: Incidence (surveillance) study for identification of new nosocomial infections for one year & 9 days from (17 of April 2007 to 26 of April 2008). The total population: All neonates attending NICU in pediatric AUH from the Obstetric Department of Maternal–Healthcare Assiut University Hospital. The target population is all neonates that developed nosocomial infections within 72 hrs of admission. The presence of risk factors was studied. A practical guide completed for each patient, which included: Demographic risk factors as age, birth weight, sex, single or multiple births, type of delivery, premature rupture of membranes, variables recorded daily as enteral feeding, parenteral feeding, mechanical ventilation and indwelling catheter. Differences between patients with and without NI for discrete variables were estimated by the chi-square test. The association between risk factors and the presence of NIs were estimated by relative risk. Results: from a total of 990 admitted neonates, the incidence rate and the incidence density rate were 16% and 24.45% infections per 1000 patient-days, respectively. Case fatality rate was 91.8%. The following risk factors were associated with NIs (P <0.05): mechanical ventilation, umbilical catheter, prematurity, birth weight less than 1500 g, use of ryle, transport outside NICU, premature rupture of membrane & peripheral vascular catheter. Conclusion: The risk of NIs increases with increasing invasive devices, decreasing birth weight, and gestational age. There is an increased mortality rate associated with NI. Strategies to minimize the impact of risk factors must be identified.