Abstract: OBJECTIVE To explore the efficacy of ATP bioluminescence assay on improving the compliance and qualifying rate of medical staff in an intensive-care unit (ICU). METHODS A total of 41 medical staff were enrolled in the study from Dec. 2013 to Mar. 2014. The baselines status hand hygiene among medical staff was investigated, then ATP bioluminescence was used to monitor and evaluate hand hygiene quality. After intervention, the hand hygiene was investigated, and compared with the baselines data. RESULTS After detection by ATP and feedback intervention, the hand hygiene compliance rate increased from 56.25% to 69.92% (P<0.05), and the qualified rate of hand hygiene increased from 71.21% to 95.77%(P<0.05). In intervention process, before and after hand-washing, the RLU values were significantly different(P<0.05). CONCLUSION The ATP bioluminescence on the detection and timely feedback can improve the compliance rate and qualified rate of hand hygiene.