Diagnosis and treatment of active tuberculosis (ATB) as well as latent tuberculosis infection (LTBI) are required for efective tuberculosis (TB) control, especially in TB endemic area. The usefulness of conventional tests to distinguish between ATB and LTBI has remained challenging. The present study was aimed to demonstrate the usefulness of the serological response to synthetic peptides from Mycobacterium tuberculosis (Mtb) antigens for discrimination between ATB and LTBI in Warao Amerindians. Serum IgG antibody levels were measured by the indirect ELISA assay using 22 designed and synthesized peptides derived from immunogenic Mtb ESAT-6 and Ag85A proteins. A total of 211 adult Warao Amerindians were included; cases with active TB (ATB, n=75), latent TB infection (LTBI, n=85) and non-infected (NI, n=51). The approach’s diagnostic information was compared using receiver operating characteristic (ROC) curves. For ATB diagnostic performance between ATB and NI; ESAT-6; P-12037 had 100% of sensitivity (AUC=0.812; 0.733 to 0.891 95% CI); and Ag85A; P-10997 had 100% of specifcity (AUC=0.691; 0.597 to 0.785 95% CI); and ATB and LTBI; Ag85A; P-29878 had 100% of sensitivity (AUC=0.741; 0.666–0.817 95% CI), and P-29879 had 99% of specifcity (AUC=0.679; 0.593–0.765 95% CI). While that ESAT-6 P-12037 also allowed diferentiation between LTBI and NI or healthy ones. It had 98.8% of sensitivity and 98.0% of specifcity (AUC=0.640; 0.545–0.735 95% CI). The potential of combination-antigen immunoassays with peptides could discriminate between Warao Amerindians with ATB, LTBI and NI. Further validation of this approach could lead to developing a complementary tool for rapid diagnosis of TB infections.