ObjectiveTo explore the effect of ATP fluorescence detection on onsite monitoring and supervision of healthcareassociated infection management . MethodsATP bioluminescence analyzer was used to detect the contamination status of hands of health care workers(HCWs), the object surfaces, and the cleaning tools in all quarters of 2015, the detection results were timely given feedback, and improvement measures were put forward. ResultsA total of 1 294 specimens were detected, the overall qualified rate was 62.75%. The qualified rates of hands of HCWs, object surfaces, and cleaning tools increased from 54.35%, 50.30%, and 60.26% in the first quarter to 76.42%, 64.80%, and 79.52% in the fourth quarter respectively，tendency chisquare test showed that difference was statistically significant (all P＜0.05). The median of relative light unit (RLU) of hands of HCWs, object surfaces, and cleaning tools were 20.00, 85.00, and 35.00, respectively. ConclusionATP fluorescence detection for onsite monitoring and supervision for cleaning and disinfection effect can promote the continuous quality improvement of hand hygiene and environmental cleanliness.