Background Healthcare workers suffered mental burden, especially in the period of COVID-19. Professional quality of life quality is suitable to measure how healthcare workers feel in medical aid team. Current evidence of impact of professional quality of life on hand hygiene behavior or even IPC measures was limited, especially in emerging infectious disease period. This study aimed to assess the prevalence of burnout, secondary traumatic stress and compassion satisfaction and explore their impact on self-reported hand hygiene behavior among medical aid team in Wuhan, China, where strict management was conducted to prevent COVID-19 spread and guarantee healthcare workers’ health. Results A cross-sectional study was conducted using online questionnaire covering professional quality of life and self-reported hand hygiene behavior based on COVID-19 guideline. A total of 1,734 healthcare workers were surveyed. The prevalence of burnout, secondary trauma and compassion satisfaction were low and average levels (69.61 and 30.39%), low and average levels (33.33 and 66.21%), average and high levels (49.65 and 49.71%), respectively. Burnout was negatively associated with overall hand hygiene (Coef. =-0.088, p<0.001), low hand hygiene (Coef. =-0.109, p<0.001), medium hand hygiene (Coef. =-0.088, p<0.001) and high hand hygiene (Coef. =-0.065, p<0.001). Conclusions Healthcare workers with higher compassion satisfaction reported higher hand hygiene compared to the lower. Healthcare workers in medical aid team experience lower level burnout, and higher level of compassion satisfaction during COVID-19 pandemic compared to the general period. The lower burnout and higher compassion satisfaction are associated with higher self-reported hand hygiene behavior. Burnout and compassion satisfaction in healthcare workers should be emphasize and need interventions targeting. The management of healthcare workers in Wuhan, China may be constructive for the future medical aid team.