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<h4>Background</h4>About 26% of the world's population may have latent tuberculosis infection (LTBI). Health care workers are a high-risk category because of their professional exposure.<h4>Methods</h4>This cross-sectional study assessed the LTBI burden among health care workers in Afghanistan, a high-TB-burden country. We selected health facilities using a systematic sampling technique and invited all workers at the targeted health facilities to participate. Participants were interviewed about sociodemographic and exposure variables and received tuberculin skin tests for LTBI.<h4>Results</h4>Of the 4,648 health care workers invited to participate, 3,686 had tuberculin skin tests. The prevalence of LTBI was found to be 47.2% (1,738 workers). Multivariate analysis showed that a body mass index of ≥ 30 and marriage were associated with an increased risk of LTBI. Underweight (body mass index of ≤ 18 and below) and normal body mass index had no association with increased risk of LTBI.<h4>Conclusion</h4>LTBI is high among health care workers in Afghanistan. We recommend instituting infection control measures in health facilities and screening workers for timely TB diagnosis.