BACKGROUND: Tuberculosis-specific ELISPOT assay (T-SPOT.TB, Oxford Immunotec, UK) is a test that detects interferon-gamma producing T-cells after stimulating patient's lymphocytes with two kinds of tuberculosis-specific antigens (ESAT-6 and CFP-10). We evaluated clinical usefulness of T-SPOT.TB test in Korean adults with intermediate burden of tuberculosis and high rate of BCG vaccination at birth. METHODS: T-SPOT.TB test was performed in 79 patients and 64 healthy volunteers and these patients and volunteers were classified into four groups: group 1, patients with active tuberculosis (n=30); group 2, patients with not-active tuberculosis (n=27); group 3, patients without tuberculosis (n=24); group 4, healthy volunteers without tuberculosis history (n=62). Positive rates and average spot counts of T-SPOT.TB test were obtained among these groups. RESULTS: Positive rates of group 1 (96.4%) and group 2 (92.3%) were higher than those of group 3 (31.6%) and group 4 (27.4%) (P<0.0001). The sensitivity deduced from group 1 and specificity deduced from group 4 of T-SPOT.TB test were 96.4% and 72.6%, respectively. The average spot counts of group 1 and 2 were higher than those of group 3 and 4 (P<0.001). There was a tendency of increasing positive rate with increasing age. Overall agreement between T-SPOT.TB test and tuberculin skin test was 63.8% (kappa=0.29). CONCLUSIONS: T-SPOT.TB test would be a very useful screening and supplementary test for diagnosis of tuberculosis due to its high sensitivity. However, positive results of T-SPOT.TB should be cautiously interpreted because of high positivity in treated tuberculosis patients and healthy volunteers in Korea.