- INTRODUCTION
- MATERIALS AND METHODS
- Chemicals and reagents.
- Leishmania promastigote secreted antigens.
- Preparation of ELISA test plates.
- Enzyme-linked immunosorbent assay (ELISA).
- Final Can-Leish IgG ELISA procedure.
- Negative control serum samples from unexposed non-endemic dogs.
- Samples from endemic dogs for evaluating Can-Leish IgG ELISA.
- Direct agglutination test (DAT).
- Immunofluorescence antibody test (IFAT).
- Specificity testing.
- Data analyses.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Alvar J, Canavate C, Molina R, Moreno J, Nieto J, 2004. Canine leishmaniasis. Adv Parasitol 57 :1–88.
-
2
Dereure J, Pratlong F, Dedet J-P, 1999. Geographical distribution and the identification of parasites causing canine leishmaniasis in the Mediterranean basin. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 18–25.
-
3
Maurício IL, Stothard JR, Miles MA, 2000. The strange case of Leishmania chagasi. Parasitol Today 16 :188–189.
-
4
Porrozi R, da Costa MVS, Teva A, Falqueto A, Ferreira AL, dos Santos CD, Fernandes AP, Gazzinelli RT, Campos-Neto A, Grimaldi G Jr, 2007. 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 14 :544–548.
-
5
Miles MA, Vexenat JA, Furtado Campos JH, Fonseca de Castro JA, 1999. Canine leishmaniasis in Latin America: control strategies for visceral leishmaniasis. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 46–53.
-
6
Castro-Ferreira E, Lana M, Carneiro M, Reis AB, Paes DV, da Silva ES, Schallig H, Gontijo CMF, 2007. Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations. Vet Parasitol 146 :235–241.
-
7
Ferrer LM, 1999. Clinical aspects of canine leishmaniasis. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 6–10.
-
8
Neogy AB, Vouldoches I, Silva OA, Tselentis Y, Lascombe JC, Segalen T, Rzepka D, Monjour L, 1992. Serodiagnosis and screening of canine visceral leishmaniasis in an endemic area of Corsica: applicability of a direct agglutination test and immunoblot analysis. Am J Trop Med Hyg 47 :772–777.
-
9
Orndorff GR, Cooper BA, Smith W, Ryan JR Jr, 2000. Canine visceral leishmaniasis in Sicily. Mil Med 165 :29–32.
-
10
Ashford DA, Bozza M, Freire M, Miranda JC, Sherlock I, Eulalio C, Lopes U, Fernandes O, Degrave W, Barker RH Jr, Badaro R, David JR, 1995. Comparison of the polymerase chain reaction and serology for the detection of canine visceral leishmaniasis. Am J Trop Med Hyg 53 :251–255.
-
11
Dietze R, Falqueto A, Valli LCP, Rodriques TP, Boulos M, Corey R, 1995. Diagnosis of canine visceral leishmaniasis with a dot-enzyme-linked immunosorbent assay. Am J Trop Med Hyg 53 :40–42.
-
12
Evans TG, Vasconcelos IAB, Lima IW, Teixeira JM, Mcullife IT, Lopes UG, Pearson RD, Vasconcelos AW, 1990. Canine visceral leishmaniasis in northeast Brazil: assessment of serodiagnostic methods. Am J Trop Med Hyg 42 :118–123.
-
13
Rajasekariah GR, Ryan JR, Hillier SR, Yi LP, Stiteler JM, Cui L, Smithyman AM, Martin SK, 2001. Optimisation of an ELISA for the serodiagnosis of visceral leishmaniasis using in vitro derived promastigote antigens. J Immunol Methods 252 :105–119.
-
14
Ryan JR, Smithyman AM, Rajasekariah G, Hochberg L, Stiteler JM, Martin SK, 2002. Enzyme-linked immunosorbent assay based on soluble promastigote antigen detects immunoglobulin M (IgM) and G (IgG) antibodies in sera from cases of visceral and cutaneous leishmaniasis. J Clin Microbiol 40 :1037–1043.
-
15
Martin SK, Thuita-Harun L, Adoyo-Adoyo M, Wasunna KM, 1998. A diagnostic ELISA for visceral leishmaniasis based on antigen from media conditioned by Leishmania donovani promastigotes. Ann Trop Med Parasitol 92 :571–577.
-
16
Harith AE, Kolk AHJ, Leeuwenburg J, Muigai R, Huigen E, Jelsma T, Kager PA, 1988. Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J Clin Microbiol 26 :1321–1325.
-
17
Sharifi I, Daneshvar H, 1996. The prevalence of visceral leishmaniasis in suspected canine reservoirs in southeastern Iran. Iran J Med Sci 21 :1–5.
-
18
Moreno J, Alvar J, 2002. Canine leishmaniasis: epidemiological risk and the experimental model. Trends Parasitol 18 :399–404.
-
19
Duprey ZH, Steurer FJ, Rooney JA, Kirchhoff LV, Jackson JE, Rowton E, Schantz PM, 2006. Canine visceral leishmaniasis, United States and Canada, 2000–2003. Emerg Infect Dis 12 :440–446.
-
20
Slappendel RJ, Teske E, 1999. A review of canine leishmaniasis outside endemic areas. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 54–59.
-
21
Teske E, van Knappen F, Beijer EGM, Slappendel RJ, 2002. Risk of infection with Leishmania spp in the canine population in the Netherlands. Acta Vet Scand 43 :195–201.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 293 | 207 | 12 |
| Full Text Views | 276 | 9 | 0 |
| PDF Downloads | 60 | 9 | 0 |
A Novel Exo-antigen-based ELISA for the Detection of Canine Leishmaniasis
G-Halli R. Rajasekariah
G-Halli R. Rajasekariah
Cellabs Pty Ltd, Australia; Department of Veterinary Sciences, CECAV, University of Trás-os-Montes e Alto Douro, Vila Real, Portugal; Walter Reed Project, USMRU-K, Kenya Medical Research Institute, Nairobi, Kenya
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Luis Cardoso
Luis Cardoso
Cellabs Pty Ltd, Australia; Department of Veterinary Sciences, CECAV, University of Trás-os-Montes e Alto Douro, Vila Real, Portugal; Walter Reed Project, USMRU-K, Kenya Medical Research Institute, Nairobi, Kenya
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Diane A. Dogcio
Diane A. Dogcio
Cellabs Pty Ltd, Australia; Department of Veterinary Sciences, CECAV, University of Trás-os-Montes e Alto Douro, Vila Real, Portugal; Walter Reed Project, USMRU-K, Kenya Medical Research Institute, Nairobi, Kenya
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Samuel K. Martin
Samuel K. Martin
Cellabs Pty Ltd, Australia; Department of Veterinary Sciences, CECAV, University of Trás-os-Montes e Alto Douro, Vila Real, Portugal; Walter Reed Project, USMRU-K, Kenya Medical Research Institute, Nairobi, Kenya
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Anthony M. Smithyman
Anthony M. Smithyman
Cellabs Pty Ltd, Australia; Department of Veterinary Sciences, CECAV, University of Trás-os-Montes e Alto Douro, Vila Real, Portugal; Walter Reed Project, USMRU-K, Kenya Medical Research Institute, Nairobi, Kenya
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Dogs which are infected with leishmania parasites serve as major reservoir hosts for zoonotic visceral leishmaniasis. The incidence of zoonotic visceral leishmaniasis is rising in many countries. This may be associated with the continuing drift of people and their pets from rural areas into peri-urban settings, particularly at the fringe of large cities. At the same time, there is evidence of adaptation of sand fly vectors to these urban settings. This has created an alarming situation because, even though domestic and stray dogs may be infected, many remain asymptomatic but are still highly infectious to the sand fly vectors and thus pose a serious threat to human health. Over half of the infected dogs have asymptomatic infections and current assays are not sensitive enough under field conditions to distinguish asymptomatic from symptomatic dogs. There is an urgent need for a specific and sensitive screening tool for use in the field. We have previously demonstrated that promastigote exo-antigen-based ELISAs can be used in the specific diagnosis of human visceral leishmaniasis (HVL). A cocktail of exo-antigens prepared from three species (L. infantum, L. donovani, and L. major) was used to develop and optimize a canine ELISA assay. Serum samples from dogs with a variety of pathological conditions but living in a non-leishmania endemic area were used as negative controls and their reactivity was used to determine a cut-off value for the ELISA. Samples from dogs residing in a leishmania endemic area were tested in parallel using direct agglutination (DAT), immunofluorescence (IFAT), and ELISA. The ELISA results correlated closely (100%) with the clinical symptoms, and were elevated in one asymptomatic dog. This sample was also found to be positive by IFAT. Based on its sensitivity and specificity, the cocktail exo-antigen-based ELISA may prove useful, even at 1:2,000 serum dilutions, for screening dogs in different geographical regions of the world.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- Chemicals and reagents.
- Leishmania promastigote secreted antigens.
- Preparation of ELISA test plates.
- Enzyme-linked immunosorbent assay (ELISA).
- Final Can-Leish IgG ELISA procedure.
- Negative control serum samples from unexposed non-endemic dogs.
- Samples from endemic dogs for evaluating Can-Leish IgG ELISA.
- Direct agglutination test (DAT).
- Immunofluorescence antibody test (IFAT).
- Specificity testing.
- Data analyses.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Alvar J, Canavate C, Molina R, Moreno J, Nieto J, 2004. Canine leishmaniasis. Adv Parasitol 57 :1–88.
-
2
Dereure J, Pratlong F, Dedet J-P, 1999. Geographical distribution and the identification of parasites causing canine leishmaniasis in the Mediterranean basin. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 18–25.
-
3
Maurício IL, Stothard JR, Miles MA, 2000. The strange case of Leishmania chagasi. Parasitol Today 16 :188–189.
-
4
Porrozi R, da Costa MVS, Teva A, Falqueto A, Ferreira AL, dos Santos CD, Fernandes AP, Gazzinelli RT, Campos-Neto A, Grimaldi G Jr, 2007. 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 14 :544–548.
-
5
Miles MA, Vexenat JA, Furtado Campos JH, Fonseca de Castro JA, 1999. Canine leishmaniasis in Latin America: control strategies for visceral leishmaniasis. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 46–53.
-
6
Castro-Ferreira E, Lana M, Carneiro M, Reis AB, Paes DV, da Silva ES, Schallig H, Gontijo CMF, 2007. Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations. Vet Parasitol 146 :235–241.
-
7
Ferrer LM, 1999. Clinical aspects of canine leishmaniasis. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 6–10.
-
8
Neogy AB, Vouldoches I, Silva OA, Tselentis Y, Lascombe JC, Segalen T, Rzepka D, Monjour L, 1992. Serodiagnosis and screening of canine visceral leishmaniasis in an endemic area of Corsica: applicability of a direct agglutination test and immunoblot analysis. Am J Trop Med Hyg 47 :772–777.
-
9
Orndorff GR, Cooper BA, Smith W, Ryan JR Jr, 2000. Canine visceral leishmaniasis in Sicily. Mil Med 165 :29–32.
-
10
Ashford DA, Bozza M, Freire M, Miranda JC, Sherlock I, Eulalio C, Lopes U, Fernandes O, Degrave W, Barker RH Jr, Badaro R, David JR, 1995. Comparison of the polymerase chain reaction and serology for the detection of canine visceral leishmaniasis. Am J Trop Med Hyg 53 :251–255.
-
11
Dietze R, Falqueto A, Valli LCP, Rodriques TP, Boulos M, Corey R, 1995. Diagnosis of canine visceral leishmaniasis with a dot-enzyme-linked immunosorbent assay. Am J Trop Med Hyg 53 :40–42.
-
12
Evans TG, Vasconcelos IAB, Lima IW, Teixeira JM, Mcullife IT, Lopes UG, Pearson RD, Vasconcelos AW, 1990. Canine visceral leishmaniasis in northeast Brazil: assessment of serodiagnostic methods. Am J Trop Med Hyg 42 :118–123.
-
13
Rajasekariah GR, Ryan JR, Hillier SR, Yi LP, Stiteler JM, Cui L, Smithyman AM, Martin SK, 2001. Optimisation of an ELISA for the serodiagnosis of visceral leishmaniasis using in vitro derived promastigote antigens. J Immunol Methods 252 :105–119.
-
14
Ryan JR, Smithyman AM, Rajasekariah G, Hochberg L, Stiteler JM, Martin SK, 2002. Enzyme-linked immunosorbent assay based on soluble promastigote antigen detects immunoglobulin M (IgM) and G (IgG) antibodies in sera from cases of visceral and cutaneous leishmaniasis. J Clin Microbiol 40 :1037–1043.
-
15
Martin SK, Thuita-Harun L, Adoyo-Adoyo M, Wasunna KM, 1998. A diagnostic ELISA for visceral leishmaniasis based on antigen from media conditioned by Leishmania donovani promastigotes. Ann Trop Med Parasitol 92 :571–577.
-
16
Harith AE, Kolk AHJ, Leeuwenburg J, Muigai R, Huigen E, Jelsma T, Kager PA, 1988. Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J Clin Microbiol 26 :1321–1325.
-
17
Sharifi I, Daneshvar H, 1996. The prevalence of visceral leishmaniasis in suspected canine reservoirs in southeastern Iran. Iran J Med Sci 21 :1–5.
-
18
Moreno J, Alvar J, 2002. Canine leishmaniasis: epidemiological risk and the experimental model. Trends Parasitol 18 :399–404.
-
19
Duprey ZH, Steurer FJ, Rooney JA, Kirchhoff LV, Jackson JE, Rowton E, Schantz PM, 2006. Canine visceral leishmaniasis, United States and Canada, 2000–2003. Emerg Infect Dis 12 :440–446.
-
20
Slappendel RJ, Teske E, 1999. A review of canine leishmaniasis outside endemic areas. In Canine Leishmaniasis: An Update. Proceedings of the International Canine Leishmaniasis Forum. Barcelona, Spain, pp. 54–59.
-
21
Teske E, van Knappen F, Beijer EGM, Slappendel RJ, 2002. Risk of infection with Leishmania spp in the canine population in the Netherlands. Acta Vet Scand 43 :195–201.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 293 | 207 | 12 |
| Full Text Views | 276 | 9 | 0 |
| PDF Downloads | 60 | 9 | 0 |