Objective To explore the method of quickly mastering the process of personal protective equipment (PPE) putting on and taking off by health care workers(HCWs), so as to provide reference for further standardized training. Methods From January to February 2020, 344 HCWs were divided into two groups and received the skill training of PPE putting on and taking off process. 209 HCWs in group 1 were from other clinical departments, 135 HCWs in group 2 were from medical team aid to Hubei, fever clinic, and isolation wards. The common training subjects of both groups were theoretical knowledge learning, video learning and manipulate training, on this basis, group 2 added simulation training examination and on-site teaching subject in isolation wards, qualified rates and learning curves of PPE putting on and taking off in two groups of HCWs were compared and analyzed. Results The qualified rate of PPE putting on and taking off in group 2 was higher than that in group 1 (90.37% vs 69.86%), difference was statistically significant (P<0.05). In group 2, the learning curve of PPE putting on and taking off process skill entered a stable period after 9 times of manipulation, correct rate of manipulation continued to rise with the increase of manipulation times; when manipulation training reached 15 times, it entered the proficiency period; in group 1, the learning curve needed 13 times of manipulation to enter the stable period, and 21 times of manipulation to enter the proficiency period. Conclusion Learning curve of PPE putting on and taking off skill of HCWs can be shortened by adding simulation training examination and on-site teaching subject in isolation wards, which is helpful for HCWs to master PPE putting on and taking off skill as soon as possible, and reduce the risk of coronavirus disease 2019 (COVID-19).