Abstract Objective A systematic review and meta-analysis was conducted to evaluate the agreement between Tuberculin Skin Test (TST) and Quantiferon (QFT) in screening for tuberculosis (TB) infection among healthcare workers (HCWs) and to estimate associations between TST and QFT agreement and variables of interest, such as Bacillus Calmette-Guérin (BCG) vaccination and incidence of TB. Methods Cross-sectional and longitudinal studies on HCWs, published in English until October 2013, comparing TST and QFT results, were selected. For each study Cohen’s κ value and a 95% confidence interval were calculated. Summary measures and indexes of heterogeneity between studies were calculated. Results 29 studies were selected comprising a total of 11,434 HCWs. Cohen’s κ for agreement between TST and QFT for 24 of them was 0.28 (95% CI 0.22 to 0.35), with the best value in high TB incidence countries and the lowest rate of BCG vaccination. Conclusion Currently, there is no gold standard for TB screening and the most-used diagnostic tools show low agreement. For evidence-based health surveillance in HCWs, occupational physicians need to consider a number of factors influencing screening results, such as TB incidence, vaccination status, age and working seniority.