Abstract: OBJECTIVE To establish a training system for infection control and prevention knowledge that is suitable for the healthcare logistics cleaning staff so as to facilitate the logistics management. METHODS A total of 129 logistics cleaning staff were recruited from Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from Nov. 2023 to Jul. 2024 and were randomly divided into two groups. The conventional training method and simulation-based training method developed in this study were employed to carry out the training of donning and doffing personal protective equipment, hand hygiene, preparation of disinfectant solutions, terminal cleaning and handling of blood and other secretions. The training effects were assessed through the uniform evaluation form after the training. RESULTS The score of disinfectant solution preparation (88.50(80.42,94.33)points, score of donning and doffing personal protective equipment 91.50(86.48,94.13)points and score of handling secretions such as blood and terminal disinfection 88.50(77.17,92.84)points of the simulation-based training group were improved after the training (P＜0.05). The female staff were superior to the male staff in standardability of disinfectant solutions preparation (P＜0.05). The logistics cleaning staff with different education backgrounds varied in the standardability of disinfectant solutions preparation(P＜0.05); the higher the education background, the lower the average score of assessment. The logistics cleaning staff with different working years varied in the standardability of disinfectant solutions preparations, disposal of secretions like blood and terminal disinfection (P＜0.05), and the staff with less than 1 year of working years were most familiar with the disinfectant solution preparations. The logistics cleaning staff were all satisfied with the simulation training method, which accordingly increased their workload or anxiety. CONCLUSIONS The assessment effect of the logistics cleaning staff is remarkably improved after they receive the specialized simulation training, and it is necessary to further boost the practical ability of the logistics cleaning staff, the newly hired staff in particularly.