MS patients are at increased risk for tuberculosis reactivation due to immunosuppressive therapies. Prevalence of latent tuberculosis in MS from our country is unknown. To assess the prevalence and determinants of latent tuberculosis infection (LTBI) in MS patients from Colombia. Multicentric, cross-sectional, retrospective study involving confirmed MS patients. LTBI was defined as a positive tuberculin skin test in asymptomatic patients with normal chest imaging. The prevalence (95 %CI) of LTBI was estimated and compared to the general prevalence with a one-sample proportion test. Determinants of LTBI were analysed using adjusted logistic regression. 481 patients were included, with a mean age of 42 years and a mean MS duration of 10 years. The prevalence of LTBI was 6.24 % (95 %CI: 4.25-8.78), significantly lower than that of the general population (21.11 %; p < 0.001). Male sex (OR: 2.52; 95 %CI: 1.00-6.28, p = 0.046) was independently associated with LTBI. LTBI is significantly less prevalent among our sample compared to the general population, and is associated with male sex.