Objective To explore the effective training approach of putting on and removing of personal protective equipment(PPE). Methods Health care workers(HCWs) were randomly divided into three groups(group 1:video training; group 2:on-site demonstration training; group 3:one-to-one practical training), training on putting on and removing of PPE was conducted, after the training, three groups of HCWs were checked, training effect was evaluated. Assessment on steps of putting on and removing of PPE was performed by table score and qualitative method (smearing fluorescence powder before removing PPE to judge whether fluorescence powder was contaminated on clothes or skin). Results 169 persons were checked in each group, qualified rate of assessment of group 3 was 85.21%, which was higher than 56.21% of group 2 and 34.32% of group 1; among three categories of posts(nursing, medical technician and doctors), the qualified rate of assessment of nursing was the highest (71.57%, 141/197), and qualified rate of medical technicians was the lowest (42.11%, 64/152); in the assessment of steps of putting on and removing of PPE, the lowest qualification rates were the tightness test of wearing medical mask as well as removing of one-piece protective clothes+hand hygiene (64.69% and 60.95%, respectively). There were statistically significant differences among three groups of trained HCWs, three different job posts, as well as steps of putting on and removing of PPE(all P<0.005). Qualified rate of fluorescence qualitative detection was 69.43%. Conclusion One-to-one training is better than on-site demonstration and video training, more attention should be paid to the tightness test of the medical protective mask during the training, when PPE is being removed, contamination on clothes or skin should be avoided.