Objective To apply infection control workshop APP-assisted concealed on-the-spot observation method in surveying hand hygiene(HH) compliance of health care workers(HCWs). Methods All HCWs in a hospital from January 2016 to October 2018 were surveyed, infection control workshop APP-assisted concealed on-the-spot observation method was adopted to survey HH compliance rates of HCWs since January 2017, difference in HH compliance rates in 2016 (adopting paper questionnaire-assisted concealed on-the-spot observation method), 2017, and January-October of 2018 were compared. Results In 2016, 2017, and January-October of 2018, 1 201, 1 237, and 1 234 HCWs were surveyed respectively, 6 539, 22 708, and 17 509 HH indications were surveyed respectively. HH compliance rate was 89.45% in 2016 revealed by paper questionnaire-assisted concealed on-the-spot observation method, 60.76% in 2017 revealed by APP-assisted concealed on-the-spot observation method, and increased to 69.06% in January-October of 2018. Compliance rates of HH among HCWs in different years were significantly different (P<0.001). Conclusion Compared with paper questionnaire-assisted concealed on-the-spot observation, infection control workshop APP-assisted concealed on-the-spot observation method can obtain more accurate data of HH compliance of HCWs, which is of guiding significance for improving HH compliance.