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      Immunodiagnosis of Canine Visceral Leishmaniasis Using Mimotope Peptides Selected from Phage Displayed Combinatorial Libraries

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          Abstract

          ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections by Trypanosoma cruzi may occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to soluble Leishmania antigen that showed cross-reactivity with anti- T. cruzi IgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL.

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          Most cited references35

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          Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface.

          G. Smith (1985)
          Foreign DNA fragments can be inserted into filamentous phage gene III to create a fusion protein with the foreign sequence in the middle. The fusion protein is incorporated into the virion, which retains infectivity and displays the foreign amino acids in immunologically accessible form. These "fusion phage" can be enriched more than 1000-fold over ordinary phage by affinity for antibody directed against the foreign sequence. Fusion phage may provide a simple way of cloning a gene when an antibody against the product of that gene is available.
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            Manual de vigilância e controle da leishmaniose visceral

            MS Brasil (2006)
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              Canine leishmaniasis: epidemiological risk and the experimental model.

              Increasing risk factors are making zoonotic visceral leishmaniasis a growing public health concern in many countries. Domestic dogs constitute the main reservoir of Leishmania infantum and Leishmania chagasi, and play a key role in the transmission to humans. New reagents and tools allow the detailed investigation of canine leishmaniasis, permitting the monitoring of the immunological status of dogs in both natural and experimental infections. Such studies are essential to determine the basis of the canine protective immune response and to establish a laboratory model, a significant aspect for the development of vaccines against canine leishmaniasis.
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                Author and article information

                Journal
                Biomed Res Int
                Biomed Res Int
                BMRI
                BioMed Research International
                Hindawi Publishing Corporation
                2314-6133
                2314-6141
                2015
                29 January 2015
                : 2015
                : 401509
                Affiliations
                1Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP 486, Belo Horizonte 31270-901, MG, Brazil
                2Departamento Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, CP 486, Belo Horizonte 31270-901, MG, Brazil
                3SysDiag CNRS-BioRad UMR 3145, Cap Delta/Parc Euromédecine, 1682 rue de la Valsière, CS 61003, 34184 Montpellier Cedex 4, France
                4Departamento de Análises Clínicas, Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto 35400-000, MG, Brazil
                5Escola de Veterinária, Universidade Federal de Minas Gerais, CP 486, Belo Horizonte 31270-901, MG, Brazil
                Author notes
                *Ricardo Toshio Fujiwara: fujiwara@ 123456icb.ufmg.br

                Academic Editor: Mehdi Chenik

                Article
                10.1155/2015/401509
                4325972
                fc23952e-ffea-4be1-9484-8e7ae96ec331
                Copyright © 2015 Christina Monerat Toledo-Machado et al.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 17 November 2014
                : 27 December 2014
                : 11 January 2015
                Categories
                Research Article

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