Chagas' disease represents an important public health problem. In Mexico most studies have been performed using Trypanosoma cruzi' antigens extracted from strains of other geographical origins. This work was aimed at developing a reactive antigen to perform serological diagnosis of Chagas' disease, using Mexican T. cruzi strains. We prepared antigenic extracts from epimastigotes, trypomastigotes and sphaeromastigotes of three Mexican strains. Parasites homogenate was obtained by lysis and sonication, solubilized proteins were analyzed by SDS-PAGE, Western-blot assays, and ELISA to determine the reactivity against sera from chagasic reference serum and chagasic and leishmaniasic patients and healthy donors. Western Blot profiles revealed, with the reference serum, eleven main components between 212 to 25 kDa; however, five bands corresponding to 74, 44, 31, 25 and 18 kDa antigens were recognized by the T. cruzi reactive sera from Mexican chagasic patients, which did not cross-react with Leishmania mexicana. Antigens from the Tequesquitengo strain yielded the best reactivity in the enzymatic immunoassay, thus enabling us to propose their use for serodiagnoses of Chagas' disease in Mexico.