ObjectiveTo evaluate the effectiveness of hospital environmental cleaning practice by fluorescence labeling method.MethodsFrom January to February 2015, 312 ward rooms in 7 hospitals were chosen, high touch object surface were labeled with fluorescence, after object surface being cleaned by cleaners, clearance rates of fluorescence labeling were calculated (as baseline survey data), training and onsite guidance for cleaners were performed(intervention measures), fluorescence labeling clearance effect before and after intervention was compared.ResultsA total of 110 ward rooms were performed baseline survey, the fluorescence labeling clearance rate of 2 856 touched clean surface was only 50.81%, the quantitative evaluation value was 45.70; after intervention, 202 rooms were surveyed, fluorescence labeling clearance rate of 3 992 touched clean surface enhanced to 79.23%, the quantitative evaluation value was 76.30；there was significant difference in fluorescence labeling clearance rate between before and after intervention （χ2=612.14，P<0.05）. In the baseline survey, the clearance rates of fluorescence labeling on touched surface of medical instruments, hospital beds, and toilets were 46.07%, 37.80%, and 25.20% respectively; after intervention , the clearance rates were 80.59%, 75.90%, and 51.70%，respectively. After intervention, fluorescence labeling clearance rates of beepers, toilet seat covers, toilet electrical switches, and chairs were low, the clearance rates of these touched surface in baseline survey were＜30%，after intervention were 47.03%-68.32%；the clearance rates of other high touch surface were all>75%. ConclusionFluorescence labeling method can directly reflect the operation quality of cleaners, and improve the cleanliness of ward environment, it is a simple, inexpensive and objective globally popular method for evaluating hospital environment cleanliness.