Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications

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Show simple item record Costa, Lourena Emanuele Lima, Mayara Ingrid Sousa Chávez-Fumagalli, Miguel Angel Menezes-Souza, Daniel Martins, Vivian Tamietti Duarte, Mariana Costa Lage, Paula Sousa Lopes, Eliane Gonçalves Paiva Lage, Daniela P. Ribeiro, Tatiana Gomes Andrade, Pedro H R De Magalhães Soares, Danielle Ferreira Soto Álvarez, Manuel Tavares, Carlos Alberto Pereira Goulart, Luiz Ricardo Coelho, Eduardo Anton̂io Ferraz
dc.contributor.other UAM. Departamento de Biología Molecular es_ES
dc.contributor.other Centro de Biología Molecular Severo Ochoa es_ES 2014-12-03T11:35:58Z 2014-12-03T11:35:58Z 2014-01-01
dc.identifier.citation Clinical and Vaccine Immunology 21.1 (2014): 96-106 en_US
dc.identifier.issn 1556-6811 (print) en
dc.identifier.issn 1556-679X (online) en
dc.description.abstract Visceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy and Trypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected with Leishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n 31) compared to those from vaccinated dogs (n 21), experimentally infected dogs with cross-reactive parasites (n 23), and healthy controls (n 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no crossreactivity with T. cruzi- or Ehrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes of L. infantum antigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs en_US
dc.description.sponsorship This work was supported by grants from the Pró-Reitoria de Pesquisa of UFMG (supported 03/2013), the Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (INCT Nano-Biofar), Rede Nanobiotec/Brasil-UFU (CAPES), PRONEX-FAPEMIG (APQ-01019- 09), FAPEMIG (APQ-00496-11 and APQ-00819-12), and CNPq (APQ- 472090/2011-9 and APQ-482976/2012-8). E.A.F.C. and L.R.G. are recipients of grants from CNPq. M.A.C.-F. is the recipient of a grant from FAPEMIG/CAPES es_ES
dc.format.extent 11 pag. es_ES
dc.format.mimetype application/pdf en
dc.language.iso eng en
dc.publisher American Society for Microbiology. en_US
dc.relation.ispartof Clinical and Vaccine Immunology en_US
dc.rights © 2014, American Society for Microbiology en_US
dc.subject.other Leishmaniasis es_ES
dc.subject.other Leishmania infantum en_US
dc.subject.other Potential serodiagnosis applications en_US
dc.title Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications en_US
dc.type article en
dc.subject.eciencia Biología y Biomedicina / Biología es_ES
dc.relation.publisherversion es_ES
dc.identifier.doi 10.1128/CVI.00583-13 es_ES
dc.identifier.publicationfirstpage 96 es_ES
dc.identifier.publicationissue 1 es_ES
dc.identifier.publicationlastpage 106 es_ES
dc.identifier.publicationvolume 21 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion en
dc.rights.accessRights openAccess en
dc.authorUAM Soto Álvarez, Manuel (260583)

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