Login Register Cart 0
Generic placeholder image

Current Pharmaceutical Biotechnology

Editor-in-Chief

ISSN (Print): 1389-2010
ISSN (Online): 1873-4316

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Diagnosis of Canine Visceral Leishmaniasis by Flow Cytometry Serology using the rMELEISH Multiepitope Antigen Coupled in a Functional Bead

Author(s): Henrique Bonifácio Moura, Henrique Gama Ker, Luana S. Ramos, Lais M. Nogueira, Juliana M. Machado, Daniel S. Dias, Rita Alaíde Leandro Rodrigues, Mariana Campos da Paz, Rafael Gonçalves Teixeira-Neto, Alexandre Barbosa Reis, Alexsandro S. Galdino* and Eduardo S. Silva*

Volume 25, Issue 17, 2024

Published on: 23 February, 2024

Page: [2290 - 2299] Pages: 10

DOI: 10.2174/0113892010268142231226115140

Price: $65

Abstract

Background: Visceral leishmaniasis (VL) is a zoonotic disease, with dogs being the main reservoir of the Leishmania infantum parasite.

Objective: To develop a new flow cytometry test to diagnosis canine VL (CVL) diagnosis.

Methods: The current study addresses a new flow cytometry test using beads coupled to the multiepitope antigen rMELEISH.

Results: In the study set of samples a sensitivity (87.1%) and specificity (89.9%) was observed. Considering the dogs’ clinical status, 20/20 (100.0%) of the symptomatic sera tested positive, while 19/22 (86.4%) of the oligosymptomatic and 16/20 (80.0%) of asymptomatic were positive. In the non-infected control, all samples (0/30) tested as negative. In the cross-reaction control, the test was more efficient in dogs infected with L. braziliensis (2/10) and Trypanosoma cruzi (0/10), than those with Babesia canis (4/10) and Ehrlichia canis (4/10). Dogs immunized with different vaccines (Leishmune, Leish-Tec®, or LBSap) did not present serological reactivity.

Conclusion: The flow cytometry serology through coupling the antigen rMELEISH in functional beads showed high accuracy in diagnosing CVL.

« Previous Next »
[1]
Baneth, G.; Koutinas, A.F.; Solano-Gallego, L.; Bourdeau, P.; Ferrer, L. Canine leishmaniosis – new concepts and insights on an expanding zoonosis: Part one. Trends Parasitol., 2008, 24(7), 324-330.
[http://dx.doi.org/10.1016/j.pt.2008.04.001] [PMID: 18514028]
[2]
Alvar, J.; Vélez, I.D.; Bern, C.; Herrero, M.; Desjeux, P.; Cano, J.; Jannin, J.; Boer, M.; Team, W.L.C. Leishmaniasis worldwide and global estimates of its incidence. PLoS One, 2012, 7(5), e35671.
[http://dx.doi.org/10.1371/journal.pone.0035671] [PMID: 22693548]
[3]
Pan American Health Organization. Leishmaniasis: Epidemiological report for the americas. PAHO, 2022, 11, 1-12.
[4]
Araújo, V.E.M.; Pinheiro, L.C.; Almeida, M.C.M.; Menezes, F.C.; Morais, M.H.F.; Reis, I.A.; Assunção, R.M.; Carneiro, M. Relative risk of visceral leishmaniasis in Brazil: A spatial analysis in urban area. PLoS Negl. Trop. Dis., 2013, 7(11), e2540.
[http://dx.doi.org/10.1371/journal.pntd.0002540] [PMID: 24244776]
[5]
Nunes, C.M.; Pires, M.M.; da Silva, K.M.; Assis, F.D.; Filho, J.G.; Perri, S.H.V. Relationship between dog culling and incidence of human visceral leishmaniasis in an endemic area. Vet. Parasitol., 2010, 170(1-2), 131-133.
[http://dx.doi.org/10.1016/j.vetpar.2010.01.044] [PMID: 20181428]
[6]
Teixeira-Neto, R.G.; da Silva, E.S.; Nascimento, R.A.; Belo, V.S.; de Oliveira, C.L.; Pinheiro, L.C.; Gontijo, C.M.F. Canine visceral leishmaniasis in an urban setting of Southeastern Brazil: An ecological study involving spatial analysis. Parasit. Vectors, 2014, 7(1), 485.
[http://dx.doi.org/10.1186/s13071-014-0485-7] [PMID: 25326767]
[7]
Coura-Vital, W.; Marques, M.J.; Veloso, V.M.; Roatt, B.M.; Aguiar-Soares, R.D.O.; Reis, L.E.S.; Braga, S.L.; Morais, M.H.F.; Reis, A.B.; Carneiro, M. Prevalence and factors associated with Leishmania infantum infection of dogs from an urban area of Brazil as identified by molecular methods. PLoS Negl. Trop. Dis., 2011, 5(8), e1291.
[http://dx.doi.org/10.1371/journal.pntd.0001291] [PMID: 21858243]
[8]
Laurenti, M.D.; Rossi, C.N.; Matta, V.L.R.; Tomokane, T.Y.; Corbett, C.E.P.; Secundino, N.F.C.; Pimenta, P.F.P.; Marcondes, M. Asymptomatic dogs are highly competent to transmit Leishmania (Leishmania) infantum chagasi to the natural vector. Vet. Parasitol., 2013, 196(3-4), 296-300.
[http://dx.doi.org/10.1016/j.vetpar.2013.03.017] [PMID: 23562649]
[9]
Romero, G.A.S.; Boelaert, M. Control of visceral leishmaniasis in latin america-a systematic review. PLoS Negl. Trop. Dis., 2010, 4(1), e584.
[http://dx.doi.org/10.1371/journal.pntd.0000584] [PMID: 20098726]
[10]
de Paiva-Cavalcanti, M.; de Morais, R.C.S.; Pessoa-e-Silva, R.; Trajano-Silva, L.A.M.; Gonçalves-de-Albuquerque, S.C.; Tavares, D.H.C.; Brelaz-de-Castro, M.C.A.; Silva, R.F.; Pereira, V.R.A. Leishmaniases diagnosis: An update on the use of immunological and molecular tools. Cell Biosci., 2015, 5(1), 31.
[http://dx.doi.org/10.1186/s13578-015-0021-2] [PMID: 26097678]
[11]
Barbosa de Castro, R.; Badaró de Moraes, J.V.; Costa Bressan, G.; de Souza Vasconcellos, R.; Silva-Júnior, A.; Lopes Rangel Fietto, J. Antigens and their diagnostic performance for Canine Visceral Leishmaniasis: A critical review. Vet. Parasitol., 2022, 301, 109638.
[http://dx.doi.org/10.1016/j.vetpar.2021.109638] [PMID: 34920304]
[12]
Soto, M.; Requena, J.M.; Quijada, L.; Alonso, C. Multicomponent chimeric antigen for serodiagnosis of canine visceral leishmaniasis. J. Clin. Microbiol., 1998, 36(1), 58-63.
[http://dx.doi.org/10.1128/JCM.36.1.58-63.1998] [PMID: 9431920]
[13]
Fonseca, A.M.; Faria, A.R.; Rodrigues, F.T.G.; Nagem, R.A.P.; Magalhães, R.D.M.; Cunha, J.L.R.; Bartholomeu, D.C.; de Andrade, H.M. Evaluation of three recombinant Leishmania infantum antigens in human and canine visceral leishmaniasis diagnosis. Acta Trop., 2014, 137, 25-30.
[http://dx.doi.org/10.1016/j.actatropica.2014.04.028] [PMID: 24801885]
[14]
Faria, A.R.; de Castro Veloso, L.; Coura-Vital, W.; Reis, A.B.; Damasceno, L.M.; Gazzinelli, R.T.; Andrade, H.M. Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. PLoS Negl. Trop. Dis., 2015, 9(1), e3429.
[http://dx.doi.org/10.1371/journal.pntd.0003429] [PMID: 25569685]
[15]
García, V.S.; Gonzalez, V.D.G.; Gugliotta, L.; Burna, A.; Demonte, A.; Arias, D.G.; Cabeza, M.S.; Guerrero, S.A. Development of a simple and economical diagnostic test for canine leishmaniasis. Exp. Parasitol., 2017, 182, 9-15.
[http://dx.doi.org/10.1016/j.exppara.2017.08.011] [PMID: 28867354]
[16]
Vale, D.L.; Lage, D.P.; Machado, A.S.; Freitas, C.S.; de Oliveira, D.; Galvani, N.C.; Fernandes, B.B.; Luiz, G.P.; Oliveira, J.S.; Oliveira-da-Silva, J.A.; Ramos, F.F.; Santos, T.T.O.; Siqueira, W.F.; Alves, L.A.; Chávez-Fumagalli, M.A.; de Magalhães-Soares, D.F.; Silveira, J.A.G.; Bueno, L.L.; Fujiwara, R.T.; Machado-de-Ávila, R.A.; Martins, V.T.; Coelho, E.A.F. Serodiagnosis of canine leishmaniasis using a novel recombinant chimeric protein constructed with distinct B-cell epitopes from antigenic Leishmania infantum proteins. Vet. Parasitol., 2021, 296, 109513.
[http://dx.doi.org/10.1016/j.vetpar.2021.109513] [PMID: 34225189]
[17]
Dias, D.S.; Machado, J.M.; Ribeiro, P.A.F.; Machado, A.S.; Ramos, F.F.; Nogueira, L.M.; Gonçalves, A.A.M.; Ramos, L.S.; Gandra, I.B.; Coutinho, F.S.; Santos, M.D.; Silva, J.O.D.; Chávez-Fumagalli, M.A.; Teixeira-Neto, R.G.; Chaves, A.T.; Campos-da-Paz, M.; Souza, A.A.; Giunchetti, R.C.; Freitas, S.M.; Lyon, S.; de Magalhães-Soares, D.F.; Silveira, J.A.G.; Silva, E.S.; Coelho, E.A.F.; Galdino, A.S. A novel recombinant multiepitope-based protein applied to the serodiagnosis of both canine and human visceral leishmaniasis. Pathogens, 2023, 12(2), 302.
[18]
Travi, B.L.; Cordeiro-da-Silva, A.; Dantas-Torres, F.; Miró, G. Canine visceral leishmaniasis: Diagnosis and management of the reservoir living among us. PLoS Negl. Trop. Dis., 2018, 12(1), e0006082.
[http://dx.doi.org/10.1371/journal.pntd.0006082] [PMID: 29324838]
[19]
de Arruda, M.M.; Figueiredo, F.B.; Cardoso, F.A.; Hiamamoto, R.M.; Brazuna, J.C.M.; de Oliveira, M.R.F.; Noronha, E.F.; Romero, G.A.S. Validity and reliability of enzyme immunoassays using Leishmania major or L. infantum antigens for the diagnosis of canine visceral leishmaniasis in Brazil. PLoS One, 2013, 8(7), e69988.
[http://dx.doi.org/10.1371/journal.pone.0069988] [PMID: 23922884]
[20]
Fraga, D.B.M.; da Silva, E.D.; Pacheco, L.V.; Borja, L.S.; de Oliveira, I.Q.; Coura-Vital, W.; Monteiro, G.R.; Oliveira, G.G.S.; Jerônimo, S.M.B.; Reis, A.B.; Veras, P.S.T. A multicentric evaluation of the recombinant Leishmania infantum antigen-based immunochromatographic assay for the serodiagnosis of canine visceral leishmaniasis. Parasit. Vectors, 2014, 7(1), 136-146.
[http://dx.doi.org/10.1186/1756-3305-7-136] [PMID: 24684857]
[21]
Figueiredo, F.B.; Vasconcelos, T.C.B.; Madeira, M.F.; Menezes, R.C.; Maia-Elkhoury, A.N.S.; Marcelino, A.P.; Werneck, G.L. Validation of the Dual-path Platform chromatographic immunoassay (DPP® CVL rapid test) for the serodiagnosis of canine visceral leishmaniasis. Mem. Inst. Oswaldo Cruz, 2018, 113(11), e180260.
[http://dx.doi.org/10.1590/0074-02760180260] [PMID: 30379198]
[22]
Souto, D.E.P.; Fonseca, A.M.; Barragan, J.T.C.; Luz, R.C.S.; Andrade, H.M.; Damos, F.S.; Kubota, L.T. SPR analysis of the interaction between a recombinant protein of unknown function in Leishmania infantum immobilised on dendrimers and antibodies of the visceral leishmaniasis: A potential use in immunodiagnosis. Biosens. Bioelectron., 2015, 70, 275-281.
[http://dx.doi.org/10.1016/j.bios.2015.03.034] [PMID: 25829285]
[23]
Ramos-Jesus, J.; Pontes-de-Carvalho, L.C.; Melo, S.M.B.; Alcântara-Neves, N.M.; Dutra, R.F. A gold nanoparticle piezoelectric immunosensor using a recombinant antigen for detecting Leishmania infantum antibodies in canine serum. Biochem. Eng. J., 2016, 110, 43-50.
[http://dx.doi.org/10.1016/j.bej.2016.01.027]
[24]
Lira, R.A.; Cavalcanti, M.P.; Nakazawa, M.; Ferreira, A.G.P.; Silva, E.D.; Abath, F.G.C.; Alves, L.C.; Souza, W.V.; Gomes, Y.M. Canine visceral leishmaniosis: A comparative analysis of the EIE-leishmaniose-visceral-canina-Bio-Manguinhos and the IFI-leishmaniose-visceral-canina-Bio-Manguinhos kits. Vet. Parasitol., 2006, 137(1-2), 11-16.
[http://dx.doi.org/10.1016/j.vetpar.2005.12.020] [PMID: 16446034]
[25]
Figueiredo, F.B.; Madeira, M.F.; Nascimento, L.D.; Abrantes, T.R.; Mouta-confort, E.; Passos, S.R.L.; Schubach, T.M.P. Canine visceral leishmaniasis: Study of methods for the detection of IgG in serum and eluate samples. Rev. Inst. Med. Trop. São Paulo, 2010, 52(4), 193-196.
[http://dx.doi.org/10.1590/S0036-46652010000400005] [PMID: 21748226]
[26]
Oliveira, E.; Saliba, J.W.; Oliveira, D.; Dias, E.S.; Paz, G.F. A prototype of the direct agglutination test kit (DAT-Canis) for the serological diagnosis of canine visceral leishmaniasis. Vet. Parasitol., 2016, 221, 9-13.
[http://dx.doi.org/10.1016/j.vetpar.2016.02.006] [PMID: 27084465]
[27]
Andrade, R.A.; Silva Araújo, M.S.; Reis, A.B.; Gontijo, C.M.F.; Vianna, L.R.; Mayrink, W.; Martins-Filho, O.A. Advances in flow cytometric serology for canine visceral leishmaniasis: Diagnostic applications when distinct clinical forms, vaccination and other canine pathogens become a challenge. Vet. Immunol. Immunopathol., 2009, 128(1-3), 79-86.
[http://dx.doi.org/10.1016/j.vetimm.2008.10.308] [PMID: 19046772]
[28]
Ker, H.G.; Coura-Vital, W.; Aguiar-Soares, R.D.O.; Roatt, B.M. das Dores Moreira, N.; Carneiro, C.M.; Machado, E.M.M.; Teixeira-Carvalho, A.; Martins-Filho, O.A.; Giunchetti, R.C.; Araújo, M.S.S.; Coelho, E.A.F.; da Silveira-Lemos, D.; Reis, A.B. Evaluation of a prototype flow cytometry test for serodiagnosis of canine visceral leishmaniasis. Clin. Vaccine Immunol., 2013, 20(12), 1792-1798.
[http://dx.doi.org/10.1128/CVI.00575-13] [PMID: 24108778]
[29]
Ker, H.G.; Coura-Vital, W.; Valadares, D.G.; Aguiar-Soares, R.D.O.; de Brito, R.C.F.; Veras, P.S.T.; Fraga, D.B.M.; Martins-Filho, O.A.; Teixeira-Carvalho, A.; Reis, A.B. Multiplex flow cytometry serology to diagnosis of canine visceral leishmaniasis. Appl. Microbiol. Biotechnol., 2019, 103(19), 8179-8190.
[http://dx.doi.org/10.1007/s00253-019-10068-x] [PMID: 31388731]
[30]
De Brito, R.C.F.; Aguiar-Soares, R.D.O.; Cardoso, J.M.O.; Coura-Vital, W.; Roatt, B.M.; Reis, A.B. Recent advances and new strategies in Leishmaniasis diagnosis. Appl. Microbiol. Biotechnol., 2020, 104(19), 8105-8116.
[http://dx.doi.org/10.1007/s00253-020-10846-y] [PMID: 32845368]
[31]
Volpini, Â.C.; Passos, V.M.A.; Oliveira, G.C.; Romanha, A.J. PCR-RFLP to identify Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis causing American cutaneous leishmaniasis. Acta Trop., 2004, 90(1), 31-37.
[http://dx.doi.org/10.1016/j.actatropica.2003.10.008] [PMID: 14739020]
[32]
de Andrade, H.M.; Reis, A.B.; dos Santos, S.L.; Volpini, Â.C.; Marques, M.J.; Romanha, A.J. Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. Vet. Parasitol., 2006, 140(3-4), 231-238.
[http://dx.doi.org/10.1016/j.vetpar.2006.03.031] [PMID: 16682124]
[33]
Reis, L.E.S.; Coura-Vital, W.; Roatt, B.M.; Bouillet, L.É.M.; Ker, H.G.; Fortes de Brito, R.C.; Resende, D.M.; Carneiro, M.; Giunchetti, R.C.; Marques, M.J.; Carneiro, C.M.; Reis, A.B. Molecular diagnosis of canine visceral leishmaniasis: A comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection. Vet. Parasitol., 2013, 197(3-4), 498-503.
[http://dx.doi.org/10.1016/j.vetpar.2013.07.006] [PMID: 23953760]
[34]
Reis, A.B.; Teixeira-Carvalho, A.; Vale, A.M.; Marques, M.J.; Giunchetti, R.C.; Mayrink, W.; Guerra, L.L.; Andrade, R.A.; Corrêa-Oliveira, R.; Martins-Filho, O.A. Isotype patterns of immunoglobulins: Hallmarks for clinical status and tissue parasite density in brazilian dogs naturally infected by Leishmania (Leishmania) chagasi. Vet. Immunol. Immunopathol., 2006, 112(3-4), 102-116.
[http://dx.doi.org/10.1016/j.vetimm.2006.02.001] [PMID: 16621021]
[35]
Mancianti, F.; Gramiccia, M.; Gradoni, L.; Pieri, S. Studies on canine leishmaniasis control. 1. Evolution of infection of different clinical forms of canine leishmaniasis following antimonial treatment. Trans. R. Soc. Trop. Med. Hyg., 1988, 82(4), 566-567.
[http://dx.doi.org/10.1016/0035-9203(88)90510-X] [PMID: 3076714]
[36]
Swets, J.A. Measuring the accuracy of diagnostic systems. Science, 1988, 240(4857), 1285-1293.
[http://dx.doi.org/10.1126/science.3287615] [PMID: 3287615]
[37]
Oliveira, G.G.S.; Magalhães, F.B.; Teixeira, M.C.A.; dos-Santos, W.L.C.; Pereira, A.M.; Gomes, Y.M.; Pinheiro, C.G.M.; Moreira, E.D.; Brito, M.E.F.; Pontes de Carvalho, L.C.; Santos, L.R.; de Melo Neto, O.P.; Nascimento, M.B.; Bedor, C.N.G.; Albuquerque, A.L. Characterization of novel Leishmania infantum recombinant proteins encoded by genes from five families with distinct capacities for serodiagnosis of canine and human visceral leishmaniasis. Am. J. Trop. Med. Hyg., 2011, 85(6), 1025-1034.
[http://dx.doi.org/10.4269/ajtmh.2011.11-0102] [PMID: 22144438]
[38]
Abánades, D.R.; Arruda, L.V.; Arruda, E.S.; Pinto, J.R.A.S.; Palma, M.S.; Aquino, D.; Caldas, A.J.; Soto, M.; Barral, A.; Barral-Netto, M. Immunodominant antigens of Leishmania chagasi associated with protection against human visceral leishmaniasis. PLoS Negl. Trop. Dis., 2012, 6(6), e1687.
[http://dx.doi.org/10.1371/journal.pntd.0001687] [PMID: 22724032]
[39]
Gama Ker, H.; Dian de Oliveira Aguiar-Soares, R.; Mendes Roatt, B. das Dores Moreira, N.; Coura-Vital, W.; Martins Carneiro, C.; Teixeira-Carvalho, A.; Assis Martins-Filho, O.; Cordeiro Giunchetti, R.; da Silveira-Lemos, D.; Barbosa Reis, A. Effect of the preservative and temperature conditions on the stability of Leishmania infantum promastigotes antigens applied in a flow cytometry diagnostic method for canine visceral leishmaniasis. Diagn. Microbiol. Infect. Dis., 2013, 76(4), 470-476.
[http://dx.doi.org/10.1016/j.diagmicrobio.2013.04.007] [PMID: 23726649]
[40]
Pinelli, E.; Killick-Kendrick, R.; Wagenaar, J.; Bernadina, W.; del Real, G.; Ruitenberg, J. Cellular and humoral immune responses in dogs experimentally and naturally infected with Leishmania infantum. Infect. Immun., 1994, 62(1), 229-235.
[http://dx.doi.org/10.1128/iai.62.1.229-235.1994] [PMID: 8262632]
[41]
Coura-Vital, W.; Marques, M.J.; Giunchetti, R.C.; Teixeira-Carvalho, A.; Moreira, N.D.; Vitoriano-Souza, J.; Vieira, P.M.; Carneiro, C.M.; Corrêa-Oliveira, R.; Martins-Filho, O.A.; Carneiro, M.; Reis, A.B. Humoral and cellular immune responses in dogs with inapparent natural Leishmania infantum infection. Vet. J., 2011, 190(2), e43-e47.
[http://dx.doi.org/10.1016/j.tvjl.2011.04.005] [PMID: 21596598]
[42]
Yamasaki, M.; Inokuma, H.; Sugimoto, C.; Shaw, S.E.; Aktas, M.; Yabsley, M.J.; Yamato, O.; Maede, Y. Comparison and phylogenetic analysis of the heat shock protein 70 gene of Babesia parasites from dogs. Vet. Parasitol., 2007, 145(3-4), 217-227.
[http://dx.doi.org/10.1016/j.vetpar.2007.01.003] [PMID: 17289267]
[43]
Schubach, E.Y.P.; Figueiredo, F.B.; Romero, G.A.S. Accuracy and reproducibility of a rapid chromatographic immunoassay for the diagnosis of canine visceral leishmaniasis in Brazil. Trans. R. Soc. Trop. Med. Hyg., 2014, 108(9), 568-574.
[http://dx.doi.org/10.1093/trstmh/tru109] [PMID: 25015665]
[44]
Laurenti, M.D.; de Santana Leandro, M.V., Jr; Tomokane, T.Y.; De Lucca, H.R.L.; Aschar, M.; Souza, C.S.F.; Silva, R.M.; Marcondes, M.; da Matta, V.L.R. Comparative evaluation of the DPP® CVL rapid test for canine serodiagnosis in area of visceral leishmaniasis. Vet. Parasitol., 2014, 205(3-4), 444-450.
[http://dx.doi.org/10.1016/j.vetpar.2014.09.002] [PMID: 25257505]

Rights & Permissions Print Cite
© 2025 Bentham Science Publishers | Privacy Policy