Use of Antigen Combinations to Address Complex Leishmania-Seropositivity Patterns in Dogs Living in Canine Leishmaniosis Endemic Regions of Portugal
Abstract
:1. Introduction
2. Materials and Methods
2.1. Samples
2.2. Direct Agglutination Test
2.3. DNA Isolation
2.4. Polymerase Chain Reactions
2.5. Antigens
2.6. Enzyme-Linked Immunosorbent Assay (ELISA)
2.7. Score System
2.8. Statistical Analysis
3. Results
3.1. Cut-Off Determination by ROC Curves
3.2. Serological Survey of PT Cohort
3.3. Score Systems Using Ratio Approaches
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Carrillo, E.; Moreno, J. Cytokine profiles in canine visceral Leishmaniasis. Vet. Immunol. Immunopathol. 2009, 128, 67–70. [Google Scholar] [CrossRef] [PubMed]
- Blavier, A.; Keroack, S.; Denerolle, P.; Goy-Thollot, I.; Chabanne, L.; Cadoré, J.L.; Bourdoiseau, G. Atypical forms of canine leishmaniosis. Vet. J. 2001, 162, 108–120. [Google Scholar] [CrossRef] [PubMed]
- Paltrinieri, S.; Solano-Gallego, L.; Fondati, A.; Lubas, G.; Gradoni, L.; Castagnaro, M.; Crotti, A.; Maroli, M.; Oliva, G.; Roura, X.; et al. Guidelines for diagnosis and clinical classification of leishmaniasis in dogs. J. Am. Vet. Med. Assoc. 2010, 236, 1184–1191. [Google Scholar] [CrossRef] [PubMed]
- Solano-Gallego, L.; Koutinas, A.; Miró, G.; Cardoso, L.; Pennisi, M.G.; Ferrer, L.; Bourdeau, P.; Oliva, G.; Baneth, G. Directions for the diagnosis, clinical staging, treatment and prevention of canine leishmaniosis. Vet. Parasitol. 2009, 165, 1–18. [Google Scholar] [CrossRef]
- Almeida, M.A.O.; Jesus, E.E.V.; Sousa-Atta, M.L.B.; Alves, L.C.; Berne, M.E.A.; Atta, A.M. Clinical and serological aspects of visceral leishmaniasis in Northeast Brazilian dogs naturally infected with Leishmania chagasi. Vet. Parasitol. 2005, 127, 227–232. [Google Scholar] [CrossRef]
- Alvar, J.; Cañavate, C.; Molina, R.; Moreno, J.; Nieto, J. Canine leishmaniasis. Adv. Parasitol. 2004, 57, 1–88. [Google Scholar] [CrossRef]
- Lachaud, L.; Chabbert, E.; Dubessay, P.; Dereure, J.; Lamothe, J.; Dedet, J.-P.; Bastien, P. Value of two PCR methods for the diagnosis of canine visceral leishmaniasis and the detection of asymptomatic carriers. Parasitology 2003, 125, 197–207. [Google Scholar] [CrossRef] [Green Version]
- Laurenti, M.D.; Rossi, C.N.; da 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, 296–300. [Google Scholar] [CrossRef]
- Miró, G.; López-Vélez, R. Clinical management of canine leishmaniosis versus human leishmaniasis due to Leishmania infantum: Putting “One Health” principles into practice. Vet. Parasitol. 2018, 254, 151–159. [Google Scholar] [CrossRef]
- Quinnell, R.J.; Courtenay, O.; Davidson, S.; Garcez, L.; Lambson, B.; Ramos, P.; Shaw, J.J.; Shaw, M.-A.; Dye, C. Detection of Leishmania infantum by PCR, serology and cellular immune response in a cohort study of Brazilian dogs. Parasitology 2001, 122, 253–261. [Google Scholar] [CrossRef]
- De Paiva-Cavalcanti, M.; De Morais, R.C.S.; Pessoa-e-Silva, R.; Trajano-Silva, L.A.M.; Goncalves-de-Albuquerque Sda, C.; Tavares Dde, H.; Brelaz-De-Castro, M.C.A.; de Freitas e Silva, R.; Pereira, V.R.A. Leishmaniases diagnosis: An update on the use of immunological and molecular tools. Cell Biosci. 2015, 5, 31. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ferreira, E.D.C.; de Lana, M.; Carneiro, M.; Reis, A.B.; Paes, D.V.; da Silva, E.S.; Schallig, H.; Gontijo, C.M.F. Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations. Vet. Parasitol. 2007, 146, 235–241. [Google Scholar] [CrossRef] [PubMed]
- Islam, M.; Ahmed, I.; Sarder, A.H.; Mirza, R.; Kimura, E.; Hashiguchi, Y.; Itoh, M.; Ekram, A.R.M.S.; Shamsuzzaman, S.M. Direct agglutination test with urine samples for the diagnosis of visceral leishmaniasis. Am. J. Trop. Med. Hyg. 2004, 70, 78–82. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Adams, E.R.; Jacquet, D.; Schoone, G.; Gidwani, K.; Boelaert, M.; Cunningham, J. Leishmaniasis direct agglutination test: Using pictorials as training materials to reduce inter-reader variability and improve accuracy. PLoS Negl. Trop. Dis. 2012, 6, e1946. [Google Scholar] [CrossRef] [PubMed]
- Cañavate, C.; Bern, C.; Chicharro, C.; Blackstock, A.J.; Alvar, J.; Herrero, M.; Aparicio, P.; Cruz, I.; Argaw, D.; Nieto, J.; et al. Evaluation of two rk39 dipstick tests, direct agglutination test, and indirect fluorescent antibody test for diagnosis of visceral leishmaniasis in a new epidemic site in highland ethiopia. Am. J. Trop. Med. Hyg. 2011, 84, 102–106. [Google Scholar] [CrossRef] [Green Version]
- da Silva, D.A.; Madeira, M.D.F.; Abrantes, T.R.; Filho, C.J.D.L.B.; Figueiredo, F.B. Assessment of serological tests for the diagnosis of canine visceral leishmaniasis. Vet. J. 2013, 195, 252–253. [Google Scholar] [CrossRef]
- Sousa, S.; Lopes, A.P.; Cardoso, L.; Silvestre, R.; Schallig, H.; Reed, S.G.; da Silva, A.C. Seroepidemiological survey of Leishmania infantum infection in dogs from northeastern Portugal. Acta Trop. 2011, 120, 82–87. [Google Scholar] [CrossRef]
- Saridomichelakis, M.N. Advances in the pathogenesis of canine leishmaniosis: Epidemiologic and diagnostic implications. Vet. Dermatol. 2009, 20, 471–489. [Google Scholar] [CrossRef]
- Silvestre, R.; Schallig, H.; Cordeiro-Da-Silva, A.; Santarém, N.; Cunha, J.; Teixeira, L. Evaluation of Leishmania species reactivity in human serologic diagnosis of leishmaniasis. Am. J. Trop. Med. Hyg. 2009, 81, 202–208. [Google Scholar] [CrossRef]
- Santarem, N.; Silvestre, R.; Cardoso, L.; Schallig, H.; Reed, S.G.; Cordeiro-Da-Silva, A. Application of an improved enzyme-linked immunosorbent assay method for serological diagnosis of canine leishmaniasis. J. Clin. Microbiol. 2010, 48, 1866–1874. [Google Scholar] [CrossRef]
- Porrozzi, R.; da Costa, M.V.S.; Teva, A.; Falqueto, A.; Ferreira, A.L.; dos Santos, C.D.; Fernandes, A.P.; Gazzinelli, R.T.; Campos-Neto, A.; Grimaldi, G. Comparative evaluation of enzyme-linked immunosorbent assays based on crude and recombinant Leishmanial antigens for serodiagnosis of symptomatic and asymptomatic Leishmania infantum visceral infections in dogs. Clin. Vaccine Immunol. 2007, 14, 544–548. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lage, D.P.; Martins, V.T.; Duarte, M.C.; Costa, L.E.; Garde, E.; Dimer, L.M.; Coelho, E.A. A new leishmania-specific hypothetical protein and its non-described specific B cell conformational epitope applied in the serodiagnosis of canine visceral Leishmaniasis. Parasitol. Res. 2016, 115, 1649–1658. [Google Scholar] [CrossRef] [PubMed]
- Santarém, N.; Sousa, S.; Amorim, C.G.; De Carvalho, N.L.; Felgueiras, Ó.; Brito, M.; Da Silva, A.C. Challenges in the serological evaluation of dogs clinically suspect for canine leishmaniasis. Sci. Rep. 2020, 10, 3099. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lima, C.; Mesquita, J.R.; Brancal, H.; Vahlenkamp, T.; Teixeira, A.R.; Cardoso, L.; Amorim, C.; Santarém, N.; DA Silva, A.C. The use of Escherichia coli total antigens as a complementary approach to address seropositivity to Leishmania antigens in canine leishmaniosis. Parasitology 2017, 144, 1384–1393. [Google Scholar] [CrossRef] [Green Version]
- Rodriguez-Cortes, A.; Ojeda, A.; Francino, O.; López-Fuertes, L.; Timón, M.; Alberola, J. Leishmania Infection: Laboratory Diagnosing in the Absence of a “Gold Standard”. Am. J. Trop. Med. Hyg. 2010, 82, 251–256. [Google Scholar] [CrossRef] [Green Version]
- Gómez-Ochoa, P.; Castillo, J.A.; Lucientes, J.; Gascón, M.; Zarate, J.J.; Arbea, J.I.; Larraga, V.; Rodriguez, C. Modified direct agglutination test for simplified serologic diagnosis of leishmaniasis. Clin. Vaccine Immunol. 2003, 10, 967–968. [Google Scholar] [CrossRef]
- Siqueira, W.F.; Viana, A.G.; Cunha, J.L.R.; Rosa, L.M.; Bueno, L.L.; Bartholomeu, D.C.; Cardoso, M.S.; Fujiwara, R.T. The increased presence of repetitive motifs in the KDDR-plus recombinant protein, a kinesin-derived antigen from Leishmania infantum, improves the diagnostic performance of serological tests for human and canine visceral leishmaniasis. PLoS Negl. Trop. Dis. 2021, 15, e0009759. [Google Scholar] [CrossRef]
- Fujimori, M.; de Almeida, A.D.B.P.F.; Barrouin-Melo, S.M.; Cortez, L.R.P.D.B.; Duthie, M.S.; Hiramoto, R.M.; de Pinho, F.A.; Reed, S.G.; Sousa, V.R.F.; Souza, N.F.; et al. Validation of ELISA with recombinant antigens in serological diagnosis of canine Leishmania infantum infection. Mem. Inst. Oswaldo Cruz. 2021, 116, e200428. [Google Scholar] [CrossRef]
- Silvestre, R.; Santarém, N.; Cunha, J.; Cardoso, L.; Nieto, J.; Carrillo, E.; Moreno, J.; Cordeiro-Da-Silva, A. Serological evaluation of experimentally infected dogs by LicTXNPx–ELISA and amastigote-flow cytometry. Vet. Parasitol. 2008, 158, 23–30. [Google Scholar] [CrossRef]
- Schallig, H.D.F.H.; Canto-Cavalheiro, M.; Da Silva, E.S. Evaluation of the direct agglutination test and the rK39 dipstick test for the sero-diagnosis of visceral Leishmaniasis. Mem. Inst. Oswaldo Cruz 2002, 97, 1015–1018. [Google Scholar] [CrossRef]
- Cruz, I.; Canavate, C.; Rubio, J.M.; Morales, M.A.; Chicharro, C.; Laguna, F.; Jiménez-Mejías, M.; Sirera, G.; Videla, S.; The Spanish HIV-Leishmania Study Group; et al. A nested polymerase chain reaction (Ln-PCR) for diagnosing and monitoring Leishmania infantum infection in patients co-infected with human immunodeficiency virus. Trans. R Soc. Trop Med. Hyg. 2022, 96, S185–S189. [Google Scholar] [CrossRef]
- Santarém, N.; Cunha, J.; Silvestre, R.; Silva, C.; Moreira, D.; Ouellette, M.; Cordeiro-Da-Silva, A. The impact of distinct culture media in Leishmania infantum biology and infectivity. Parasitology 2014, 141, 192–205. [Google Scholar] [CrossRef] [PubMed]
- Castro, H.; Sousa, C.; Novais, M.; Santos, M.; Budde, H.; Cordeiro-Da-Silva, A.; Flohé, L.; Tomás, A.M. Two linked genes of Leishmania infantum encode tryparedoxins localised to cytosol and mitochondrion. Mol. Biochem. Parasitol. 2004, 136, 137–147. [Google Scholar] [CrossRef] [PubMed]
- Parikh, R.; Mathai, A.; Parikh, S.; Sekhar, G.C.; Thomas, R. Understanding and using sensitivity, specificity and predictive values. Indian J. Ophthalmol. 2008, 56, 45–50. [Google Scholar] [CrossRef]
- Chen, T.; Liu, Y.; Huang, L. ImageGP: An easy-to-use data visualization web server for scientific researchers. iMeta 2022, 1, e5. [Google Scholar] [CrossRef]
- Software, Giacomo Scarpellini—IDoStatistics, Cohen’s kappa free calculator. Available online: https://idostatistics.com/cohen-kappa-free-calculator/#calculator (accessed on 12 January 2022).
- Lima, C.; Santarém, N.; Nieto, J.; Moreno, J.; Carrillo, E.; Bartholomeu, D.C.; Bueno, L.L.; Fujiwara, R.; Amorim, C.; Cordeiro-Da-Silva, A. The use of specific serological biomarkers to detect Canileish vaccination in dogs. Front. Vet. Sci. 2019, 6, 373. [Google Scholar] [CrossRef] [Green Version]
- Cardoso, L.; Schallig, H.D.; Neto, F.; Kroon, N.; Rodrigues, M. Serological survey of Leishmania infection in dogs from the municipality of Peso da Régua (Alto Douro, Portugal) using the direct agglutination test (DAT) and fast agglutination screening test (FAST). Acta Trop. 2004, 91, 95–100. [Google Scholar] [CrossRef]
- Cortes, S.; Afonso, M.O.; Alves-Pires, C.; Campino, L. Stray dogs and leishmaniasis in urban areas, Portugal. Emerg. Infect. Dis. 2007, 13, 1431–1432. [Google Scholar] [CrossRef]
- Venturin, G.L.; Bragato, J.P.; Silva, K.L.O.; De Lima, V.M.F. Recombinant K28 antigen in ELISA in the diagnosis of canine visceral leishmaniosis. Parasite Immunol. 2015, 37, 670–673. [Google Scholar] [CrossRef]
- Lauricella, M.A.; Negri, V.; Sinagra, A.J.; Romagosa, C.M.; Benedetti, R.; Laucella, S.; Tartaglino, L.; Reed, S.G.; Luna, C.; Frias, V.F.; et al. An rK28-based immunoenzymatic assay for the diagnosis of canine visceral leishmaniasis in Latin America. Am. J. Trop. Med. Hyg. 2016, 95, 92–98. [Google Scholar] [CrossRef]
- Santarém, N.; Racine, G.; Silvestre, R.; Cordeiro-Da-Silva, A.; Ouellette, M. Exoproteome dynamics in Leishmania infantum. J. Proteom. 2013, 84, 106–118. [Google Scholar] [CrossRef] [PubMed]
- Gomes, Y.; Cavalcanti, M.P.; Lira, R.; Abath, F.; Alves, L. Diagnosis of canine visceral Leishmaniasis: Biotechnological advances. Vet. J. 2008, 175, 45–52. [Google Scholar] [CrossRef] [PubMed]
A | B | C | D | E | ||
---|---|---|---|---|---|---|
B1 | B2 | |||||
CS | ≥1 | Ø | Ø | Ø | Ø | ≥1 |
DAT | + | + | + | − | Ø | Ø |
PCR | +/− | + | − | + | Ø | Ø |
(%) | 3.3 (13/390) | 1.3 (5/390) | 5.9 (23/390) | 3.3 (13/390) | 71.5 (279/390) | 14.6 (57/390) |
SPLA | rLicTXNPx | rK39 | rK28 | rKDDR | LAM | |
---|---|---|---|---|---|---|
AUC | 1 | 0.9840 | 1 | 1 | 1 | 0.9997 |
(95% CI) | (1.000 to 1.000) | (0.9685 to 0.9996) | (1.000 to 1.000) | (1.000 to 1.000) | (1.000 to 1.000) | (0.9988 to 1.000) |
p value | <0.0001 | <0.0001 | <0.0001 | <0.0001 | <0.0001 | <0.0001 |
Cut-off | 0.075 | 0.040 | 0.127 | 0.175 | 0.098 | 0.131 |
Sensitivity (%) | 100 | 94.21 | 100 | 100 | 100 | 99.17 |
(95% CI) | (97.00 to 100) | (88.44 to 97.64) | (97.00 to 100) | (97.00 to 100) | (97.00 to 100) | (95.48 to 99.98) |
Specificity (%) | 100 | 93.10 | 100 | 100 | 100 | 100 |
(95% CI) | (88.06 to 100) | (77.23 to 99.15) | (88.06 to 100) | (88.06 to 100) | (88.06 to 100) | (88.06 to 100) |
Groups | Characteristic | Antigens | |||||||
---|---|---|---|---|---|---|---|---|---|
CS | DAT | PCR | SPLA | LicTXNPx | rK39 | rK28 | rKDDR | LAM | |
PT | 21.5 (84/390) | 22.3 (87/390) | 16.2 (63/390) | 17.7 (69/390) | 15.6 (61/390) | 15.6 (61/390) | |||
A | + | + | +/− | 100 (13/13) | 100 (13/13) | 100 (13/13) | 100 (13/13) | 100 (13/13) | 100 (13/13) |
B | Ø | + | +/− | 82.1 (23/28) | 64.3 (18/28) | 75.0 (21/28) | 85.7 (24/28) | 75.0 (21/28) | 75.0 (21/28) |
B1 | Ø | + | + | 100 (5/5) | 60.0 (3/5) | 100 (5/5) | 100 (5/5) | 100 (5/5) | 100 (5/5) |
B2 | Ø | + | Ø | 78.3 (18/23) | 65.2 (15/23) | 69.6 (16/23) | 82.6 (19/23) | 69.6 (16/23) | 69.6 (16/23) |
C | Ø | Ø | + | 20.1 (3/13) | 38.5 (5/13) | 20.1 (3/13) | 15.4 (2/13) | 30.80 (4/13) | 20.1 (3/13) |
D | Ø | Ø | Ø | 10.8 (30/279) | 15.4 (43/279) | 5.4 (15/279) | 6.4 (18/279) | 4.3 (12/279) | 5.4 (15/279) |
E | + | Ø | Ø | 26.3 (15/57) | 14.0 (8/57) | 19.3 (11/57) | 21.0 (12/57) | 19.3 (11/57) | 15.8 (9/57) |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Lima, C.S.; Esteves, S.; Costa, I.; Brancal, H.; Lima, C.; Amorim, C.; Cardoso, L.; Santarém, N.; Cordeiro-da-Silva, A. Use of Antigen Combinations to Address Complex Leishmania-Seropositivity Patterns in Dogs Living in Canine Leishmaniosis Endemic Regions of Portugal. Microorganisms 2022, 10, 2018. https://doi.org/10.3390/microorganisms10102018
Lima CS, Esteves S, Costa I, Brancal H, Lima C, Amorim C, Cardoso L, Santarém N, Cordeiro-da-Silva A. Use of Antigen Combinations to Address Complex Leishmania-Seropositivity Patterns in Dogs Living in Canine Leishmaniosis Endemic Regions of Portugal. Microorganisms. 2022; 10(10):2018. https://doi.org/10.3390/microorganisms10102018
Chicago/Turabian StyleLima, Carla Silva, Sofia Esteves, Inês Costa, Hugo Brancal, Clara Lima, Célia Amorim, Luís Cardoso, Nuno Santarém, and Anabela Cordeiro-da-Silva. 2022. "Use of Antigen Combinations to Address Complex Leishmania-Seropositivity Patterns in Dogs Living in Canine Leishmaniosis Endemic Regions of Portugal" Microorganisms 10, no. 10: 2018. https://doi.org/10.3390/microorganisms10102018