ObjectiveTo investigate the occurrence of healthcareassociated infection(HAI) in different weight groups of neonates in neonatal intensive care unit (NICU) and general neonatal wards, and find out the high risk factors for HAI.MethodsTargeted monitoring on hospitalized neonates in the neonatal department of a hospital in 2016 and JanuaryNovember 2017 was prospectively surveyed, HAI rate, device usage rate and deviceassociated infection rate in different weight groups of neonates in NICU and general neonatal wards were compared.ResultsFrom 2016 to November 2017, a total of 3 872 neonates were monitored, the total hospitalization days were 42 427 days, 56 patients had 62 cases of infection, HAI rate and HAI case rate were 1.45% and 1.60% respectively. HAI rate and HAI case rate in NICU were 5.28% and 6.03% respectively；HAI rate and HAI case rate in general neonatal wards were both 0.46%, constituent ratio of septicemia ranked first of infection in both NICU and general neonatal wards, accounting for 43.75% and 50.00% respectively, followed by lower respiratory tract(LRT) infection, accounting for 27.08% and 21.43% respectively. HAI rate of different weight groups of neonates in NICU and general wards were both significantly different（χ2=107.78, 46.65，respectively, both P＜0.0001), the lower weight of the neonates, the higher HAI rate; ventilator usage rate in neonates was positively correlated with HAI(r=0.973, P=0.027), the higher device usage rate, the higher HAI rate. Fungal septicemia rate per 1 000 days in rainy season was higher than that of nonrainy season (0.53‰ vs 0.37‰). A total of 70 specimens were detected, 45 strains of bacteria and fungi were isolated, pathogens were all from LRT and blood system, Pseudomonas aeruginosa was the main pathogen from LRT, Klebsiella pneumoniae and Candida krusei were the main pathogens from blood system.ConclusionTargeted monitoring on neonatal HAI can timely find out high risk factors for neonatal HAI, analyze the causes of infection, formulate corresponding prevention and control measures, reduce HAI rate, and ensure the medical safety of neonates.