Purpose: The purpose of this study was to provide basic data on the infection prevention management program, which is one of the infectious disease control program by identifying the prevalence and risk factors of latent tuberculosis infection (LTBI) in healthcare workers. Methods: We surveyed a total of 3,046 LTBI test results, including those of 2,269 existing staff and 777 new employees. An interferon-gamma release assay (IGRA) for the diagnosis of LTBI was performed using QuantiFERON？？-TB Gold In-Tube (QFT-IT). The risk factors of LTBI were analyzed using logistic regression analysis. Results: The overall prevalence of LTBI was 16.0% (487/3,046). The prevalence of LTBI in the existing staff was 17.9% (406/2,269) and the prevalence of LTBI in new employees was 10.4% (81/777). Multivariate logistic regression analysis revealed that the risk factors of latent tuberculosis infection among the existing staff were gender, age and work period wheres, the risk factor amongst the new employees depended on their age. Conclusion: The LTBI was not related to the type of occupation and work unit. Therefore, while establishing an infection control program for the prevention of tuberculosis infection at medical institurions, institutional heads and infection control experts should encompass a policy for all the employees.