Tuberculosis screening is recommended for all health care workers. We evaluated the prevalence of latent tuberculosis infection among 939 hospital workers of Tor Vergata University teaching hospital in Rome, Italy, in the period 2007-2013, by using the QuantiFERON Gold In-Tube (QFT) test. The mean age of subjects tested was 31 years. The prevalence of positive subjects (cut-off 0.35 UI/ml) was 5.5% (46/939) and the mean age of those who tested positive was 39 years. The low rate of positivity may be partly related to the higher reliability of QFT in comparison to tuberculin skin testing.