This study investigates the identification of differentially recognized T cell epitopes in *Mycobacterium tuberculosis* (Mtb) antigens during the spectrum of tuberculosis (TB) infection. A proteome-wide screen of 20,610 Mtb-derived peptides in 21 patients with active TB (ATB) revealed 137 unique epitopes, 16% of which were recognized by two or more participants. These epitopes were predominantly derived from cell wall and cell processes antigens. The study also found differential recognition of antigens between ATB participants and interferon-gamma release assay (IGRA) positive/negative individuals. An ATB-specific peptide pool (ATB116) was developed, consisting of epitopes exclusively recognized by ATB participants, which could distinguish patients with pulmonary ATB from IGRA +/− individuals with a sensitivity of over 60% and a specificity exceeding 80%. This comprehensive screen identified infection stage-specific epitopes and antigens that could be useful for diagnostics and measuring Mtb-specific immune responses. The study highlights the importance of understanding the complexity of T cell responses during natural infections and the potential of ATB116 as a diagnostic tool.This study investigates the identification of differentially recognized T cell epitopes in *Mycobacterium tuberculosis* (Mtb) antigens during the spectrum of tuberculosis (TB) infection. A proteome-wide screen of 20,610 Mtb-derived peptides in 21 patients with active TB (ATB) revealed 137 unique epitopes, 16% of which were recognized by two or more participants. These epitopes were predominantly derived from cell wall and cell processes antigens. The study also found differential recognition of antigens between ATB participants and interferon-gamma release assay (IGRA) positive/negative individuals. An ATB-specific peptide pool (ATB116) was developed, consisting of epitopes exclusively recognized by ATB participants, which could distinguish patients with pulmonary ATB from IGRA +/− individuals with a sensitivity of over 60% and a specificity exceeding 80%. This comprehensive screen identified infection stage-specific epitopes and antigens that could be useful for diagnostics and measuring Mtb-specific immune responses. The study highlights the importance of understanding the complexity of T cell responses during natural infections and the potential of ATB116 as a diagnostic tool.