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Background: Health care workers are at risk for Mycobacterium tuberculosis (MTB) infection., Objectives: To perform an occupational health survey among 621 employees of a 800-bed third level care hospital covered by MTB surveillance., Methods: Statistical analysis was applied to results from tuberculin skin test (TST), QuantiFERON - TB Gold in tube assay (QFT), PPD-ELISA for serum antibodies, and occupational or vaccine data., Results: 29.1% of subjects were TST positive, 18.5% were QFT positive. In 23% of subjects no correlation between these tests was found, presumably linked to BCG-vaccination, since TST positivity was 4 times higher among vaccinated subjects, whereas both tests correlated well in unvaccinated subjects. QFT values above 2 IU/ml were significantly associated with positive TST and age over 40 years. Working in MTB risk level 4 was significantly associated with QFT, TST and PPD-antibody levels, suggesting booster effects by repeated exposure. No clear correlation was observed with medical specializations but significantly higher QFTpositivity was found in subjects not assigned to the classical medical professions and originating from MTB high risk areas., Conclusions: These results shift the focus on maintenance personnel, who mostly worked in MTB risk level 2 areas. The less positive QFT results in vaccinated subjects highlight QFT's advantage as a screening tool and argue for a protective effect of the BCG-vaccine, although percentages of vaccinated persons varied largely between different medical professions. Interestingly, the percentage of QFT positive persons was lower among subjects reporting MTB exposure than those who were not aware of exposure events.