Next-generation ELISA diagnostic assay for Chagas Disease based on the combination of short peptidic epitopes.
Mucci, J., Carmona, S. J., Volcovich, R., Altcheh, J., Bracamonte, E., Marco, J. D., Nielsen, M., Buscaglia, C. A. and Aguero, F.
Instituto de Investigaciones Biotecnologicas (IIB)-Instituto Tecnologico de Chascomus (INTECH), Universidad de San Martin (UNSAM)-Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), San Martin, Buenos Aires, Argentina.
Servicio de Parasitologia y Chagas, Hospital de Ninos Ricardo Gutierrez, Ciudad Autonoma de Buenos Aires, Argentina.
Instituto de Patologia Experimental, Facultad de Ciencias de la Salud, Universidad Nacional de Salta (UNSa)-Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), Salta, Argentina.
Department of Bio and Health Informatics, Technical University of Denmark, DK Lyngby, Denmark.
Chagas Disease, caused by the protozoan Trypanosoma cruzi, is a major health and economic problem in Latin America for which no vaccine or appropriate drugs for large-scale public health interventions are yet available. Accurate diagnosis is essential for the early identification and follow up of vector-borne cases and to prevent transmission of the disease by way of blood transfusions and organ transplantation. Diagnosis is routinely performed using serological methods, some of which require the production of parasite lysates, parasite antigenic fractions or purified recombinant antigens. Although available serological tests give satisfactory results, the production of reliable reagents remains laborious and expensive. Short peptides spanning linear B-cell epitopes have proven ideal serodiagnostic reagents in a wide range of diseases. Recently, we have conducted a large-scale screening of T. cruzi linear B-cell epitopes using high-density peptbide chips, leading to the identification of several hundred novel sequence signatures v associated to chronic Chagas Disease. Here, we performed a serological assessment of 27 selected epitopes and of their use in a novel multipeptide-based diagnostic method. A combination of 7 of these peptides were finally evaluated in ELISA format against a panel of 199 sera samples (Chagas-positive and negative, including sera from Leishmaniasis subjects). The multipeptide formulation displayed a high diagnostic performance, with a sensitivity of 96.3% and a specificity of 99.15%. Therefore, the use of synthetic peptides as diagnostic tools are an attractive alternative in Chagas' disease diagnosis.
PLoS Neglected Tropical Diseases 11(10): e0005972 (2017)