Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/30409
Title: Development of a fluorescent based immunosensor for the serodiagnosis of canine Leishmaniasis combining immunomagnetic separation and flow cytometry
Authors: Sousa, Susana
Cardoso, Luıs
Reed, Steven G.
Reis, Alexandre Barbosa
Martins Filho, Olindo Assis
Silvestre, Ricardo
Silva, Anabela Cordeiro da
Affilliation: Universidade do Porto. Instituto de Biologia Molecular e Celular. Parasite Disease Group. Porto, Portugal
Universidade do Porto. Instituto de Biologia Molecular e Celular. Parasite Disease Group. Porto, Portugal/Universidade de Tras-os-Montes e Alto Douro. Departamento de Ciencias Veterinarias. Vila Real, Portugal
Infectious Disease Research Institute. Seattle, Washington, United States of America
Universidade Federal de Ouro Preto. Nucleo de Pesquisas em Ciencias Biologicas. Laboratorio de Imunopatologia. Ouro Preto, MG, Brazil/Universidade Federal de Ouro Preto. Escola de Farmacia. Departamento de Analises Clınicas. Ouro Preto, MG, Brazil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Belo Horizonte, MG, Brazil
Universidade do Porto. Instituto de Biologia Molecular e Celular. Parasite Disease Group. Porto, Portugal/Instituto Superior de Ciencias da Saude - Norte, CESPU, CRL. Departamento de Ciencias. Gandra, Portugal
Universidade do Porto. Instituto de Biologia Molecular e Celular. Parasite Disease Group. Porto, Portugal/Universidade do Porto. Faculdade de Farmacia. Departamento de Ciencias Biologicas. Porto, Portugal
Abstract: An accurate diagnosis is essential for the control of infectious diseases. In the search for effective and efficient tests, biosensors have increasingly been exploited for the development of new and highly sensitive diagnostic methods. Here, we describe a new fluorescent based immunosensor comprising magnetic polymer microspheres coated with recombinant antigens to improve the detection of specific antibodies generated during an infectious disease. As a challenging model, we used canine leishmaniasis due to the unsatisfactory sensitivity associated with the detection of infection in asymptomatic animals where the levels of pathogen-specific antibodies are scarce. Methodology: Ni-NTA magnetic microspheres with 1,7 mm and 8,07 mm were coated with the Leishmania recombinant proteins LicTXNPx and rK39, respectively. A mixture of equal proportions of both recombinant protein-coated microspheres was used to recognize and specifically bind anti-rK39 and anti-LicTNXPx antibodies present in serum samples of infected dogs. The microspheres were recovered by magnetic separation and the percentage of fluorescent positive microspheres was quantified by flow cytometry. Principal Findings: A clinical evaluation carried out with 129 dog serum samples using the antigen combination demonstrated a sensitivity of 98,8% with a specificity of 94,4%. rK39 antigen alone demonstrated a higher sensitivity for symptomatic dogs (96,9%), while LicTXNPx antigen showed a higher sensitivity for asymptomatic (94,4%). Conclusions: Overall, our results demonstrated the potential of a magnetic microsphere associated flow cytometry methodology as a viable tool for highly sensitive laboratorial serodiagnosis of both clinical and subclinical forms of canine leishmaniasis.
Keywords: Flow cytometry
Canine leishmaniasis
keywords: citometria de fluxo
leishmaniose canina
Issue Date: 2013
Publisher: Public Library of Science
Citation: SOUSA, Susana et al. Development of a fluorescent based immunosensor for the serodiagnosis of canine Leishmaniasis combining immunomagnetic separation and flow cytometry. PLoS Neglected Tropical Diseases, v. 7, p. e2371, 2013.
DOI: 10.1371/journal.pntd.0002371
ISSN: 1935-2735
Copyright: open access
Appears in Collections:MG - IRR - Artigos de Periódicos

Files in This Item:
File Description SizeFormat 
Development of a fluorescent based immunosensor .pdf3.78 MBAdobe PDFView/Open


FacebookTwitterDeliciousLinkedInGoogle BookmarksBibTex Format mendeley Endnote DiggMySpace

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.