Chagas disease-specific antigens: characterization of epitopes in CRA/FRA by synthetic peptide mapping and evaluation by ELISA-peptide assay

dc.creatorBottino, Carolina G.
dc.creatorGomes, Luciano Pinho
dc.creatorPereira, José B.
dc.creatorCoura, José Rodrigues
dc.creatorProvance Jr., David William
dc.creatorDe Simone, Salvatore Giovanni
dc.date2015-08-19T13:49:29Z
dc.date2015-08-19T13:49:29Z
dc.date2013
dc.date.accessioned2023-09-26T23:04:20Z
dc.date.available2023-09-26T23:04:20Z
dc.identifierBOTTINO, Carolina G.; et al. Chagas disease-specific antigens: characterization of epitopes in CRA/FRA by synthetic peptide mapping and evaluation by ELISA-peptide assay. BMC Infectious Diseases, v.13:568, 10p, 2013.
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/11536
dc.identifier10.1186/1471-2334-13-568
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8886507
dc.descriptionBackground: The identification of epitopes in proteins recognized by medically relevant antibodies is useful for the development of peptide-based diagnostics and vaccines. In this study, epitopes in the cytoplasmic repetitive antigen (CRA) and flagellar repetitive antigen (FRA) proteins from Trypanosoma cruzi were identified using synthetic peptide techniques and pooled sera from Chagasic patients. The epitopes were further assayed with an ELISA assay based on synthetic peptides. Methods: Twenty-two overlapping synthetic peptides representing the coding sequence of the T. cruzi CRA and FRA proteins were assessed by a Spot-synthesis array analysis using sera donated by patients with Chagas disease. Shorter peptides were selected that represented the determined epitopes and synthesized by solid phase synthesis to evaluate the patterns of cross-reactivities and discrimination through an ELISA-diagnostic assay. Results: The peptide Spot-synthesis array successfully identified two IgG antigenic determinants in the CRA protein and four in FRA. Bioinformatics suggested that the CRA antigens were unique to T. cruzi while the FRA antigen showed similarity with sequences present within various proteins from Leishmania sp. Subsequently, shorter peptides representing the CRA-1, CRA-2 and FRA-1 epitopes were synthesized by solid phase synthesis and assayed by an ELISA-diagnostic assay. The CRA antigens gave a high discrimination between Chagasic, Leishmaniasis and T. cruzi-uninfected serum. A sensitivity and specificity of 100% was calculated for CRA. While the FRA antigen showed a slightly lower sensitivity (91.6%), its specificity was only 60%. Conclusions: The epitopes recognized by human anti-T. cruzi antibodies have been precisely located in two biomarkers of T. cruzi, CRA and FRA. The results from screening a panel of patient sera through an ELISA assay based on peptides representing these epitopes strongly suggest that the sequences from CRA would be useful for the development of diagnostic reagents that could improve upon the sensitivity and specificity of currently available diagnostic tests. Overall, the results provide further evidence of the usefulness of identifying specific linear B-cell epitopes for improving diagnostic tools.
dc.formatapplication/pdf
dc.languageeng
dc.publisherBioMed Central
dc.rightsopen access
dc.subjectChagas Disease
dc.subjectTrypanosoma cruzi
dc.subjectCytoplasmic repetitive antigen
dc.subjectFlagellar repetitive antigen
dc.subjectEpitopes
dc.subjectSpot-synthesis
dc.subjectPeptide-ELISA
dc.subjectDoença de Chagas
dc.subjectTrypanosoma cruzi
dc.subjectEpitopos
dc.subjectEnsaio de Imunoadsorção Enzimática
dc.subjectPeptídeos
dc.titleChagas disease-specific antigens: characterization of epitopes in CRA/FRA by synthetic peptide mapping and evaluation by ELISA-peptide assay
dc.typeArticle


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