Abstract: OBJECTIVE To investigate the cognitive status of the knowledge on tuberculosis-related prevention and treatment among medical staff in a hospital and its influencing factors. METHODS A questionnaire on knowledge related to prevention and treatment of tuberculosis infection was developed, and the registered nurses and medical practitioners who have worked in the hospital for more than one year were enrolled from Jan. 2015 to Oct. 2016. The basic data of the research objects, the scores of the items in the questionnaire and the total score were analyzed. The scores of the questionnaires were analyzed by univariate analysis, and the items with statistical significance in the univariate analysis were assigned as independent variables, and the scores of the questionnaires were set as dependent variables for logistic multivariate analysis. RESULTS For the 348 respondents who completed the questionnaire, the total score was (29.84±3.45) points. The univariate analysis showed a s significant difference in the total scores of the questionnaires completed by respondents with different ages, titles, qualifications, length of service, department, experience in treatment or care of tuberculosis patients (P<0.05). The older the respondents, the higher the professional titles and academic qualifications, the longer the working years, and the more the experience in treatment or care for tuberculosis, the higher the total score of the questionnaire was. Subjects in the respiratory and infectious disease departments had higher scores than those in other departments. Multivariate logistic regression analysis showed that the professional title, education, seniority, department, and experience in treatment or care of tuberculosis patients were the influencing factors of the questionnaire score. CONCLUSION The overall cognitive level of the prevention and treatment knowledge relevant to tuberculosis among medical staff in the hospital needs to be improved, and training, physical examination and protection should be strengthened.