Abstract: OBJECTIVE To prospectively evaluate the effect of improving the quality of environmental cleaning reducing the and disinfection on incidence of septicemia in neonates. METHODS The effects of cleaning the high-frequency contact objects surfaces in neonatal wards of a three-A hospital and the intervention measures were evaluated by means of fluorescent labeling method, training and feedback, as well as intensive terminal disinfection. RESULTS The clearance rates of fluorescent markers for high-frequency contact points of the bed surfaces and instrument surfaces were significantly increased from 87.36% and 75.81% before the intervention to 99.43% and 96.62% after the intervention(χ²=2117.04,783.118; P＜0.001).The incidence of septicemia and daily mortality rate were respectively 3.44% and 2.83‰ in the neonatal wards after the intervention measures were taken, significantly lower than 5.97% and 4.62‰ before the intervention measures were taken(P＜0.05). CONCLUSION It is an effective way to conduct the monitoring and feedback by using fluorescent labeling method, carry out the education and training for cleaning staff and strengthen the terminal disinfection so as to improve the effect of environmental cleaning and disinfection and prevent the septicemia.