Purpose: The purpose of this study was to investigate knowledge, awareness, and compliance on tuberculosis prevention behavior among medical technicians and nursing assistants working in a hospital department with an increased risk of TB. Method: From February 11 to February 28, 2019, 149 medical technicians and nursing assistants working in departments at high risk of TB in a hospital in Seoul, Korea, were surveyed. Result: Knowledge score for TB was 7.66 on average for out of 10 points, awareness and compliance score for TB prevention was 37.56, and 36.86 for out of 40 points respectively. Analysis of the correlation between awareness of prevention behavior and compliance with that behavior showed a significant positive relationship. In multiple regression analysis, factors influencing TB prevention behavior were occupation ( p=.032) and compliance of tuberculosis prevention behavior (p <.001). The result was 34.1% explanatory power. Conclusion: In order to enhance the compliance of TB prevention behavior, it is necessary to develop active public relations and education programs that recognize the importance of employees themselves and bring about change in behavior, as opposed to educational approaches that convey simple knowledge.