ObjectiveTo investigate the efficacy of ATP bioluminescence assay on improving the compliance and qualifying rate of hand hygiene of rotating doctors in an intensive care unit.MethodsBaseline status of hand hygiene of doctors were investigated firstly,then the frequency and methods of monitor according to the results were made, hand hygiene were monitored by ATP bioluminescence assay, results were performed feedback, the status of hand hygiene after intervention were investigated.ResultsA total of 120 doctors participated in the baseline investigation, the total qualified rate of hand hygiene was 73.33%, hand hygiene compliance rate was only 14.17%. After field detection by ATP and feedback intervention, the qualified rate of hand hygiene increased to 82.24%（χ2=13.68，P=0.008）; handwashing compliance rate increased most during the intervention process (69.44%).ConclusionATP bioluminescence assay on the field detection and timely feedback of doctors’ hand hygiene status can improve the hand hygiene compliance and qualified rate.