Background: Nosocomial infections constitute one of the leading causes of morbidity and mortality in premature neonates in Neonatal Intensive Care Units (NICUs) and affect the duration of their hospitalization, as well as the quality of their care. Aim: The study was carried out in order to record and describe the risk factors for nosocomial infections in neonates hospitalized in NICUs. Method and Material: In this prospective cross-sectional survey, all the neonates (100%) who were admitted in the NICUs of a General Pediatric (during 7 months) and a Maternity Hospital (6 months) (n=474), constituted the population of the study. Data was collected by means of a record card including data about Demographics, Consumption of antibiotics, Infections’ surveillance, Clinical Identification and Laboratory confirmations. Analyses were conducted using SPSS statistical software (version 18.0). Results: 301 neonates (63,5%) were premature. 40,9% of the neonates developed sepsis and 34,9% primary bacteremia. Neonates hospitalized in open NICU were more likely to develop nosocomial infections compared with those who were hospitalized in closed NICU (OR 3.37, 95% CI 1.94 to 5.86, p<0.001). Premature neonates were more likely to develop nosocomial infections compared with those with a normal duration of gestational age (OR 4.46, 95% CI 2.04 to 9.72, p<0.001). Proportionally, when the duration of the intravenous therapy (OR 1.14, 95% CI 1.10 to 1.19. p <0.001) and of the hospitalization were prolonged the likelihood of nosocomial infection increased. Conclusion: Factors such as low birth weight, prematurity, intravenous therapy and mechanical ventilation were associated with nosocomial infections. Limited use of invasive methods and devices and adherence to all the principles and procedures of aseptic techniques could reduce the incidence of nosocomial infections.