Author | Faria, Angélica Rosa | |
Author | Andrade, Hélida Monteiro de | |
Author | Franco, Miriam Maria Silva Costa | |
Author | Giusta, Mário S. | |
Author | Penido, Marcus L. O. | |
Author | Grimaldi Filho, Gabriel | |
Author | Gazzinelli, Ricardo Tostes | |
Access date | 2014-10-28T11:49:44Z | |
Available date | 2014-10-28T11:49:44Z | |
Document date | 2011 | |
Citation | FARIA, Angélica et al. High-throughput analysis of synthetic peptides for the immunodiagnosis of canine visceral leishmaniasis. Plos Neglect Trop D, 5(9): e1310. | pt_BR |
ISSN | 1935-2735 | |
URI | https://www.arca.fiocruz.br/handle/icict/8680 | |
Language | eng | pt_BR |
Publisher | Public Library of Science | pt_BR |
Rights | open access | pt_BR |
Title | High-throughput analysis of synthetic peptides for the immunodiagnosis of canine visceral leishmaniasis | pt_BR |
Type | Article | pt_BR |
DOI | 10.1371/journal.pntd.0001310 | |
Abstract | Background: Visceral leishmaniasis is the most severe form of leishmaniasis. Approximately 20% of zoonotic human visceral leishmaniasis worldwide is caused by Leishmania infantum, which is also known as Leishmania chagasi in Latin America, and disease incidence is increasing in urban and peri-urban areas of the tropics. In this form of disease, dogs are the main reservoirs. Diagnostic methods used to identify Leishmania infected animals are not able to detect all of the infected ones, which can compromise the effectiveness of disease control. Therefore, to contribute to the improvement of diagnostic methods for canine visceral leishmaniasis (CVL), we aimed to identify and test novel antigens using high-throughput analysis.
Methodology/Principal Findings: Immunodominant proteins from L. infantum were mapped in silico to predict B cell epitopes, and the 360 predicted peptides were synthesized on cellulose membranes. Immunoassays were used to select the most reactive peptides, which were then investigated with canine sera. Next, the 10 most reactive peptides were synthesized using solid phase peptide synthesis protocol and tested using ELISA. The sensitivity and specificity of these peptides were also compared to the EIE-LVC Bio-Manguinhos kit, which is recommended by the Brazilian Ministry of Health for use in leishmaniasis control programs. The sensitivity and specificity of the selected synthesized peptides was as high as 88.70% and 95.00%, respectively, whereas the EIE-LVC kit had a sensitivity of 13.08% and 100.00% of specificity. Although the tests based on synthetic peptides were able to diagnose up to 94.80% of asymptomatic dogs with leishmaniasis, the EIE-LVC kit failed to detect the disease in any of the infected asymptomatic dogs.
Conclusions/Significance: Our study shows that ELISA using synthetic peptides is a technique with great potential for diagnosing CVL; furthermore, the use of these peptides in other diagnostic methodologies, such as immunochromatographic tests, could be beneficial to CVL control programs. | pt_BR |
Affilliation | Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil | pt_BR |
Affilliation | Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil | pt_BR |
Affilliation | Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brasil | pt_BR |
Affilliation | Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brasil | pt_BR |
Affilliation | Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brasil | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, MG, Brasil | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil/University of Massachusetts Medical School. Division of Infectious Diseases and Immunology Worcester, MA, United States of America/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brasil | pt_BR |
Subject | Diagnostic medicine | pt_BR |
Subject | Enzyme-linked immunoassays | pt_BR |
Subject | Leishmaniasis | pt_BR |
Subject | Parasitic diseases | pt_BR |
Subject | Peptide mapping | pt_BR |
Subject | Peptide synthesis | pt_BR |