ObjectiveTo improve hand hygiene (HH) compliance of health care workers (HCWs) in neonatal intensive care units(NICUs) in China through a series of intervention measures.MethodsA multicenter study was conducted, 17 tertiary first class hospitals in 9 provinces and cities were selected, HH compliance of HCWs in these hospitals were investigated every month according to HH investigation method of World Health Organization.From October 1 ，2013 to March 31，2014 was preintervention stage； from April 1, 2014 to September 30,2014 was postintervention stage, timely feedback and intensified training were conducted at postintervention stage. ResultsHH compliance rates of HCWs before and after intervention were 80.29% and 80.85% respectively, there was no significant difference (P＞0.05). HH compliance rates of  HCWs in different sizes of ICUs were significantly different before and after intervention (all P<0.05), in NICUs with<20 beds and  20-30 beds after the intervention were both significantly higher than before intervention (both P<0.05), while ＞30 beds were significantly lower than before intervention(P<0.001); HH compliance rate of cleaners increased from 58.82% before intervention to 68.09% after intervention (P<0.05); Of different hand hygiene indications, except before clean/aseptic task, compliance to the other HH indications were significantly different between before and after intervention(all P<0.05).ConclusionHH compliance is high among HCWs in NICUs in China, intervention measures, such as intensified training and timely feedback have certain influence in compliance to HH among HCWs at different sizes of ICUs, of different occupations, and at different HH indications.