The detection of Leishmania spp. through a disposable surface plasmon resonance (SPR) biochip, using mono- and polychromatic sources, is presented here. A highly specific and sensitive-constrained synthetic peptide (LC2) derived from a mimotope of Leishmania chagasi antigen was immobilized on the sensing region using a simple protocol. ELISA- and PCR-confirmed negative and positive sera of Leishmania chagasi infected patients were used. Detection of positive IgG showed a fast response with good sensitivity and specificity for the proposed method.