Objective To understand Hawthorne effect on compliance rate of hand hygiene(HH) of staff of diffe-rent occupations, different departments and five HH opportunities, so as to provide basis for rational use of Hawthorne effect and improve HH compliance rate. Methods The direct observation method was adopted to investigate HH compliance of the same kind of population in different departments through routine survey method and concealed survey method, results were analyzed, the consumption of hand sanitizer and hand disinfectant used at the same period was investigated. Results Consumption of HH products in routine survey group and concealed survey group were 26.9 mL/bed-day and 27.5 mL/bed-day respectively, there was no significant difference between two groups (P>0.05). HH compliance rates in routine survey group and concealed survey group were 79.7% (985/1 236) and 49.8% (646/1 298) respectively, difference was statistically significant (χ²=247.166, P<0.01). Nurses had the highest HH compliance rate, HH compliance rate of doctors in concealed survey group and routine survey group were 31.6% and 82.6% respectively, routine survey group increased by 161.4%. HH compliance rates were the highest in routine survey group before aseptic manipulation and in internal medicine group (86.5% and 88.7% respectively), HH compliance rates of concealed survey group after touching patient's blood and body fluid were both higher (69.2% and 78.5% respectively). Compared with concealed survey group, HH compliance rates before touching patients in routine survey group increased by 127.0% (81.5% and 35.9% respectively). Except for touching patients' blood and body fluid, there were significant differences in compliance rates of the other four HH indicators and different departments between two groups (all P<0.05). Conclusion Concealed survey combined with the consumption of hand sanitizer and hand disinfectant can truly reflect HH compliance rate of staff in different departments and staff with different occupations, Hawthorne effect on HH compliance in routine survey group is higher, continuous stimulation through Hawthorne effect on practical work can be used to improve HH compliance rate of health care workers.