Abstract: Objective To study the risk factors of ventilator-associated pneumonia (VAP) in newborn and the profiles of isolated pathogens. Methods The clinical data of 179 neonates in the previous group (from December 2005 to December 2008) and 331 neonates in the present group (from January 2009 to January 2012) admitted into the neonatal intensive care unit (NICU) and received mechanical ventilation for equal or longer than 48 hours were respectively reviewed and analyzed,and their isolated pathogen profile and drug sensitivity were also compared in two groups. Results The incidence of VAP in the previous group and the present group were 22.35% and 20.24% respectively, no significant difference between two groups (P>0.05) was observed. Gram-negative bacteria was the main pathogens (93.40%), the first 5 common pathogens in the previous group were Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli and Enterobacter aerogenes. However, the first 5 of isolated gram-negative pathogens in the present group were somewhat different from the previous group, which were Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter cloacae, Escherichia coli and Enterobacter aerogenes. The drug sensitivity test showed that these pathogens had a general decline in sensitivities to commonly used antibiotics, of which the carbapenem antibiotic drug sensitivity decreased significantly, there was statistically significant difference between the previous group and the present group (P<0.05). The risk factors of VAP include the duration of mechanical ventilation (OR=5.680, 95%CI:2.867-11.253), the frequency of tracheal intubation (OR=2.219, 95%CI:1.037-4.748), birth weight (OR=2.127, 95%CI:1.067-4.238) and neonatal asphyxia (OR=2.025, 95%CI:1.079-3.799) as well as the duration of hospital stay (OR=2.012, 95%CI:1.215-3.967). Conclusions According to the data in our hospital, the main pathogenic bacteria of VAP were gram-negative pathogens, which showed a general decline in sensitivities to commonly used antibiotics. The incidence of VAP was closely related to the duration of mechanical ventilation, the frequency of intubation, birth weight and the hospitalization as well as the neonatal asphyxia.