AIM: This study aims to determine the prevalence of latent tuberculosis infection (LTBI) in patients with moderate to severe rheumatoid arthritis (RA) who receive conventional and anti-cytokine therapy, to identify possible risk factors for tuberculosis (TB) and to evaluate the prophylactic treatment in positively screened patients. PATIENT AND METHODS: We conducted an observational, retrospective study in patients with RA, who underwent LTBI screening (chest X-ray, tuberculin skin test and interferon gamma release assay test - IGRA test). RESULTS: Out of 124 patients included, 7.25% of patients were diagnosed with LTBI during the treatment with conventional synthetic anti-rheumatic drugs in combination with glucocorticoids before initiation of anti-cytokine therapy. Another 21.77% were diagnosed during treatment with biologics or Janus kinase inhibitors. We confirmed the highest incidence of LTBI in TNF-treated group (66.66% LTBI positive patients), but also found positive screening in patients treated with other modalities. The mean LTBI detection time since the initiation of anti-cytokine therapy was 39.5 months (12-134 months). Active TB with clinical manifestation has occurred in one patient. Statistical analysis did not show an association between risk of LTBI and age, sex or treatment modality. CONCLUSION: The results of this study confirm the necessity of LTBI screening and long-term monitoring in RA patients treated with any kind of anti-cytokine therapy. The currently used national recommendations are sufficiently sensitive to identify TB in these patients. There remains a question of screening and prophylactic antituberculosis therapy in patients treated with conventional synthetic anti-rheumatic drugs in combination with glucocorticoids.