ObjectiveTo investigate cleanliness of hospital environmental object surfaces and hands of health care workers(HCWs).MethodsThe adenosine triphosphate (ATP) bioluminescence assay was used to detect object surfaces and hands of HCWs in a hospital, onthespot intervention was conducted.ResultsThe qualified rates of hospital environmental object surfaces and ventilatorrelevant object surfaces were 58.14% (200/344) and 69.88% (116/166) respectively, the qualified rate of ventilator tracheal intubation site was low (29.17%)；the qualified rate of telephone surfaces was the lowest (27.27%）. The qualified rates of  ventilatorrelevant object surfaces used continuously for ≥48h and <48 h were 56.70%（55/97）and  88.41%（61/69） respectively，there was significant difference between the two（χ2=19.26，P<0.01）.The qualified rates of HCWs’ hands before and after intervention were 34.18% and  85.58% respectively, relative light unit (RLU) values were （1 033.46±106.20）and （80.46±10.68）respectively，the qualified rates and RLU before and after intervention were both significantly different （both P<0.01）.ConclusionContamination of object surfaces and hands’ of HCWs in hospital dynamic environment is serious, ATP bioluminescence detection and onthespot intervention is helpful for improving cleanliness of hospital environment object surfaces and HCWs’  compliance to hand hygiene.