Abstract: OBJECTIVE To understand the status of acquisition of knowledge of infection control and observe the effect of training and influencing factors in Huangshi, Hubei province during prevention and control of COVID-19.METHODS The medical staff from all of the medical institutions of Huangshi City were enrolled in the study, the online training and check was conducted by using ‘four-step’ method from Mar 11, 2020 to Mar 24, 2020.RESULTS The coverage rate of training and assessment of medical staff was 93.48% in the city, and the average score has increased from（69.11±15.27） points in the first stage to（80.51±15.38） points in the fourth stage. The average scores of assessment of the medical staff from different areas, with different genders, age, professional backgrounds and professional titles were significantly higher in the fourth stage than in the first stage, and there（P＜0.001）. The correct rate of the knowledge points was increased from 69.17% in the first stage to 80.40% in the fourth stage, and there was significant difference（P＜0.001）. Linear regression analysis showed that the scores of the medical staff were affected by the area, gender, age, working years, professional background and title in the fourth stage.CONCLUSION The medical staff of the city are less aware of the knowledge of infection control, the status of overall mastery is remarkably improved after systematic training. The status of mastery of COVID-19-related knowledge is good, and it is necessary to focus on the training of knowledge of disinfection and sterilization.