Early detection of latent tuberculosis infection (LTBI) might prevent active TB development in healthcare workers (HCWs). The aim of the study is to assess the prevalence of LTBI among HCWs exposed to active TB, compare QFT-GIT and TST in the diagnosis of LTBI, and explore possible risk factors of LTBI.This was a cross-sectional study for a period of 6 months among 153 HCWs in high-risk departments dealing with TB infection - Beni-Suef University Hospital, Egypt.HCWs were asked to fill a questionnaire for possible LTBI risk factors, and tuberculin skin test (TST) and serum QuantiFERON test were used for LTBI screening. (-18) was used for data analysis; qualitative data were compared using Chi-square test, while associations between risk factors for TB and positive QFT or TST were analyzed by a logistic regression model.LTBI detected by QuantiFERON-TB Gold In-Tube Test (QFT-GIT) and by TST was 9.1% and 34.6%, respectively (kappa = 0.028). Logistic regression showed that departments, duration of work, the use of N95 masks, and training in infection control practices were significant predictors for positive QFT-GIT among participants ( < 0.05).Work duration of >10 years, nurse profession, diabetics, and smokers were at increased risk of having LTBI. Increased training programs and implementation of infection control measures TB to reduce the risk of LTBI are recommended.