Device-associated infections are common in Neonatal Intensive Care Units (NICUs) in accordance with the frequent use of invasive devices, and they must be continuously and closely monitored for infection control. Six hundred newborn infants hospitalized longer than 72 hours in Ege University Children's Hospital NICU between January 2008 and December 2010 were prospectively followed for occurrence of device-associated infections (central venous catheter- and umbilical catheter-associated blood stream infections [CVC/UC BSI] and ventilator-associated pneumonia [VAP]). In a total of 10,052 patient days, the VAP rate was 13.76/1000 ventilator days with a ventilator utilization ratio of 0.29, and the CVC/UC BSI rate was 3.8/1000 catheter days with a catheter utilization ratio of 0.24. The CVC/UC BSI rate was lower than national averages, being close to rates reported from developed countries. The VAP rate was higher than the national and international rates and was associated with prolonged mechanical ventilation and very low birth weight. VAP also appeared to be an important risk factor for mortality. The most frequent agents were gram-negative pathogens for VAP and coagulase-negative staphylococci for CVC/UC BSIs, with resistance patterns similar to the previous years. In conclusion, with device utilization rates similar to those in developed countries, our CVC/UC BSI rate was comparable, but the VAP rate was higher than that of the developed countries. Necessary precautions are urgently needed to decrease VAP rates and VAP-related mortality.