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Comparative Evaluation of the Antibody-detection ELISA Technique Using Microplates Precoated with Denatured Crude Antigens from Trypanosoma congolense or Trypanosoma vivax

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Abstract

Two FAO/IAEA indirect enzyme-linked immunosorbent assays (ELISA), which use microplates precoated with denatured crude Trypanosoma congolense or Trypanosoma vivax antigen for detecting anti-trypanosomal antibodies in bovine sera, were evaluated for their sensitivity, specificity and positive and negative predictive values, using 320 Ugandan field samples (known negative sera, n = 80; known positive sera, n = 80; cattle herds where control of tsetse and trypanosomosis was practised, n = 80; and cattle herds where there was no such control, n = 80). Cut-off points of 30% and 25% positivity were determined for the T. congolense and T. vivax assays, respectively, using a modified ROC (receiver operating characteristic) analysis. The T. congolense assay had estimated diagnostic sensitivity and specificity of 63.7% and 57.5%, respectively, while the T. vivax assay had estimated diagnostic sensitivity and specificity of 81.3% and 81.3%, respectively. The two assays conducted in parallel had estimated diagnostic sensitivity and specificity of 82.5% and 88.7%, respectively. Using the sera from the cattle in the area with control (detected prevalence of trypanosomosis 0%), both the T. congolense and T. vivax assays had negative and positive predictive values of 100% and 0%, respectively. Using the sera from the cattle in the area without control (detected prevalence of trypanosomosis 15%), the T. congolense assay had negative and positive predictive values of 91% and 33%, respectively, and the T. vivax assay had negative and positive predictive values of 93% and 27%, respectively. The T. congolense assay was in fair agreement with the buffy coat technique (BCT) (κ = 0.25), while the T. vivax assay was in substantial agreement with the BCT (κ = 0.625), and both assays conducted in parallel were in substantial agreement with the BCT (κ = 0.708). Both assays were found to be proficient and suitable for the diagnosis of bovine trypanosomosis, especially when used in parallel.

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REFERENCES

  • Crowther, J.R., 1995. ELISA: Theory and practice. In: J.M. Walter (ed.), Methods in Molecular Biology, vol. 42, Humana Press, Totowa, NJ, USA

    Google Scholar 

  • Desquesnes, M., 1999. Sensitivity, specificity and cross reactions in Trypanosoma spp. antibody-ELISA. Integrated Control of Pathogenic Trypanosomes and their Vectors (ICPTV), Newsletter, No. 1, September

  • Dwinger, R.H., Grieve, A.S., Snow, W.F., Rawlings, P., Jabing, B. and Williams, D.J.L., 1992. Maternal antibodies in N'dama calves kept under natural trypanosomiasis risk in The Gambia. Parasite Immunology, 14, 351-354

    Google Scholar 

  • Eisler, M.C., Lessard, P., Masake, R.A., Moloo, S.K. and Peregrine, A.S., 1998. Sensitivity and specificity of antigen-capture ELISAs for diagnosis of Trypanosoma congolense and Trypanosoma vivax infections in cattle. Veterinary Parasitology, 79, 187-201

    Google Scholar 

  • FAO/IAEA, 1998a. Trypanosomosis indirect enzyme immunoassay for detection of bovine serum antibody to Trypanosoma congolense. Joint FAO/IAEA Programme, Animal Production and Health. Bench Protocol, version-iTAB (TcAGd) Prototype 1.0

  • FAO/IAEA, 1998b. Trypanosomosis indirect enzyme immunoassay for detection of bovine serum antibody to Trypanosoma vivax. Joint FAO/IAEA Programme, Animal Production and Health. Bench Protocol, version-iTAB (TcAGd) Prototype 1.0

  • Greiner, M., Sohr, D. and Göbel, P., 1995. A modified ROC analysis for the selection of cut-off values and the definition of intermediate results of serodiagnostic tests. Journal of Immunological Methods, 85, 123-132

    Google Scholar 

  • Greiner, M., Bhat, T.S., Patzelt, R.J., Kakaire, D., Schares, G., Dietz, E., Böhning, D., Zessin, K.H. and Mehlitz, D., 1997. Impact of biological factors on the interpretation of bovine trypanosomosis serology. Preventive Veterinary Medicine, 30, 61-73

    Google Scholar 

  • Luckins, A.G., 1977. Detection of antibodies in trypanosome-infected cattle by means of a microplate enzyme-linked immunosorbent assay. Tropical Animal Health and Production, 9, 53-62

    Google Scholar 

  • Magona, J.W., Okuna, N.M., Katabazi, B., Okoth, J.O., Omollo, P., Mayende, J.S.P. and Drabile, D.C., 1998. Control of tsetse and animal trypanosomosis by combination of pour-on, trapping and chemotherapy along the Uganda-Kenya border. Revue d'élevage et de médecine vétérinaire des pays tropicaux, 51, 311-315

    Google Scholar 

  • Murray, M., Murray, P.K. and McIntyre, W.I.M., 1977. An improved parasitological technique for the diagnosis of African trypanosomiasis. Transactions of the Royal Society of Tropical Medicine and Hygiene, 7, 325-326

    Google Scholar 

  • Nantulya, V.M. and Lindqvist, K.J., 1992. Application of a monoclonal antibody-based antigen detection enzyme-linked immunosorbent assay (antigen ELISA) for field diagnosis of bovine trypanosomiasis in Nguruman, Kenya. Annals of Tropical Medicine and Parasitology, 86, 225-230

    Google Scholar 

  • Ndyabahinduka, D.C.K., 1993. Uganda Country Report. The 22nd International Scientific Council for Trypanosomosis Research and Control, Kampala, Uganda

  • OIE, 1996. Manual of standards for diagnostic tests and vaccines. Principles of validation of diagnostic assays for infectious diseases. Office International des Epizooties, 8-15

  • Okuna, N.M. and Magona, J.W., 1997. Monitoring of tsetse and trypanosomosis control programmes in South-East Uganda. International Atomic Energy Agency — TECDOC, 925, 81-85

    Google Scholar 

  • Okuna, N.M., Katabazi, B., Omollo, J.P., Magona, J. and Mayende, J.S.P., 1996. The current status and epidemiological pattern of animal trypanosomosis in different agroecological zones and farming systems in Uganda. Livestock Health Research Institute Annual Report, 1996, 21-23

  • Paris, J., Murray, M. and McOdimba, F., 1982. A comparative evaluation of the parasitological techniques currently available for the diagnosis of African trypanosomiasis in cattle. Acta Tropica, 39, 30

    Google Scholar 

  • Politzar, H., 1974. Serological studies on cattle experimentally infected with several species of African trypanosomes. Tropenmedizin und Parasitologie, 25, 22-27

    Google Scholar 

  • Rebeski, D.E., Winger, E.M., Robinson, M.M., Crowther, J.R. and Dwinger, R.H., 1999a. Current status of improved enzyme-linked immunoassay methods for diagnosis and control of African trypanosomosis conducted under the FAO/IAEA Coordinated Research Programme 3.13.D 3.20.13. Integrated Control of Pathogenic Trypanosomes and their Vectors (ICPTV) Newsletter, No. 1, September

  • Rebeski, D.E., Winger, E.M., Rogovic, B., Robinson, M.M., Crowther, J.R. and Dwinger, R.H., 1999b. Improved methods for the diagnosis of African trypanosomosis. Memoir Instituto Oswaldo Cruz, Rio de Janeiro, 94, 249-253

    Google Scholar 

  • Thrusfield, M., 1995. Veterinary Epidemiology, (Blackwell Science, Oxford)

    Google Scholar 

  • Vos, G.J. and Gardiner, P.R., 1990. Parasite-specific antibody response of ruminants infected with Trypanosoma vivax. Parasitology, 100, 93-100

    Google Scholar 

  • Wilson, A.J. and Cunningham, M.P., 1971. Immunological aspects of bovine trypanosomiasis: IV. Patterns in the production of common antibodies. Tropical Animal Health and Production, 3, 133-139

    Google Scholar 

  • Wright, P.F., Nilsson, E., Van Rooij, E.M.A., Lelenta, M. and Jeggo, M.H., 1993. Standardisation and validation of enzyme-linked immunosorbent assay techniques for the detection of antibody in infectious disease diagnosis. Revue Scientifique et Technique de l'Office International des Epizooties, 12, 435-450

    Google Scholar 

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Magona, J., Mayende, J. & Walubengo, J. Comparative Evaluation of the Antibody-detection ELISA Technique Using Microplates Precoated with Denatured Crude Antigens from Trypanosoma congolense or Trypanosoma vivax . Tropical Animal Health and Production 34, 295–308 (2002). https://doi.org/10.1023/A:1015630701923

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