Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-23T12:12:47.209Z Has data issue: false hasContentIssue false
  • English
  • Français

Detection of circulating trypanosomal antigens by double antibody ELISA using antibodies to procyclic trypanosomes

Published online by Cambridge University Press:  06 April 2009

M. K. Liu  and
T. W. Pearson
M. K. Liu
Affiliation:
Department of Biochemistry and Microbiology, University of Victoria, Victoria, B.C., CanadaV8W 2Y2
T. W. Pearson*
Affiliation:
Department of Biochemistry and Microbiology, University of Victoria, Victoria, B.C., CanadaV8W 2Y2
*
*Reprint requests to Dr T. W. Pearson.
Get access Rights & Permissions [Opens in a new window]

Summary

A double antibody sandwich ELISA technique has been developed for detection of antigens of African trypanosomes present in the sera of infected mammals. The assay uses a high titre, high affinity rabbit antiserum made to purified membranes of procyclic trypanosomes as ‘capture’ reagent and a mixture of three biotin-labelled trypanosome-specific monoclonal antibodies as detecting reagent. The monoclonal antibodies were chosen on the basis of their specificity for surface membrane antigens of Trypanosoma brucei spp., the relative abundance and solubility of their specific antigen in aqueous solvents (including serum), and the fact that each monoclonal antibody binds to distinct epitopes on the same antigen molecule. Thus, antigen capture from serum and subsequent detection was achieved using as little as 10 ng/ml of whole trypanosome lysate, or the equivalent of 5000 trypanosomes/ml when solubilized material was added to normal serum in an artificial system. Using the optimized assay, antigen was detected in the sera of trypanosome-infected mice as early as 2 days after infection with T. b. rhodesiense. The results indicate that the assay allows detection of low concentrations of specific membrane antigens of T. brucei spp. of African trypanosomes and thus may have immunodiagnostic utility.

Type
Research Article
Information
Parasitology , Volume 95 , Issue 2 , October 1987 , pp. 277 - 290
Copyright
Copyright © Cambridge University Press 1987

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Ádám, É., Nász, I., Lengyel, A., Erdei, J. & Fachet, J. (1986). Multiple copies of identical epitopes on the adenovirus hexon. Molecular Immunology 23, 755–9.CrossRefGoogle ScholarPubMed
Aiyedum, B. A. & Amedu, A. A. (1973). The capillary tube agglutination (CA) test. A proposed aid for the diagnosis of trypanosomiasis in the field. Tropenmedizin und Parasitologie 24, 67–71.Google Scholar
Aiyedum, B. A., Amedu, A. A. & Bidwell, D. E. (1976). Parallel evaluation of serological tests applied in African trypanosomiasis: a WHO collaborative study. Bulletin of the World Health Organization 54, 141–7.Google Scholar
Brun, R. & Schöenberger, M. (1979). Cultivation and in vitro cloning of procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Acta Tropica 36, 289–92.Google Scholar
DeRaadt, P. (1983). Sleeping sickness control: a new elan. In Symposium on the Diagnosis of African Sleeping Sickness due to T. gambiense, Anvers-Antwerp, (ed. Crooy, P. J.), pp. 17–20. Belgium: Smith Kline-RIT Biological Division.Google Scholar
Focus (1985). Bethesda Research Laboratories Technical Information, vol. 7, no. 4, p. 12.Google Scholar
Greenwood, F. C., Hunter, W. M. & Glover, J. S. (1963). The preparation of 131I-labelled human growth hormone of high specific radioactivity. The Biochemical Journal 89, 114–23.CrossRefGoogle Scholar
Hall, T. & Esser, K. (1984). Topologic mapping of protective and nonprotective epitopes on the variant surface glycoprotein of the WRATat 1 clone of Trypanosoma brucei rhodesiense. Journal of Immunology 132, 2059–63.CrossRefGoogle ScholarPubMed
Hill, G. C., Shimer, S. P., Caughey, B. & Sauer, L. S. (1978). Growth of infective forms of Trypanosoma rhodesiense in vitro, the causative agent of African trypanosomiasis. Science 202, 763–5.CrossRefGoogle ScholarPubMed
Hoogenraad, N., Helman, T. & Hoogenraad, J. (1983). The effect of pre-injection of mice with pristane on ascites tumor formation and monoclonal antibody production. Journal of Immunological Methods 61, 317–20.CrossRefGoogle Scholar
James, M. A., Kakoma, I., Ristic, M. & Cagnard, M. (1985). Induction of protective immunity to Plasmodium falciparium in Saimiri sciureus monkeys with partially purified exoantigens. Infection and Immunity 49, 476–80.CrossRefGoogle Scholar
Lanham, S. M. & Godfrey, D. G. (1970). Isolation of salivarian trypanosomes from man and other mammals using DEAE-cellulose. Experimental Parasitology 28, 521–34.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951). Protein measurement with folin phenol reagent. Journal of Biological Chemistry 193, 265–72.CrossRefGoogle ScholarPubMed
Luckins, A. G., Gray, A. R. & Rae, P. F. (1978). Comparison of the diagnostic value of serum immunoglobulin levels, an enzyme-linked immunosorbent assay and a fluorescent antibody test in experimental infection with Trypanosoma evansi in rabbits. Annals of Tropical Medicine and Parasitology 72, 429–41.CrossRefGoogle Scholar
Magnus, E., Van Meirvenne, N., Vervoort, T., Le Ray, D. & Wery, M. (1978). Use of freeze-dried trypanosomes in the indirect fluorescent antibody test for serodiagnosis of sleeping sickness. Annales de la Société Belge de Médecine Tropicale 58, 169–76.Google ScholarPubMed
Magnus, E., Vervoort, T. & Van Meirvenne, N. (1978). A card- agglutination test with stained trypanosomes (CATT) for the serological diagnosis of T. b. gambiense trypanosomiasis. Annales de la Société Belge de Médecine Tropicale 58, 177–85.Google ScholarPubMed
Mancini, P. R., Strickler, J. E. & Patton, C. C. (1982). Identification and partial characterization of plasma membrane polypeptides of Trypanosoma brucei. Biochimica et Biophysica Acta 688, 399–406.CrossRefGoogle ScholarPubMed
Paris, J., Murray, M. & Agure, R. (1980). An evaluation of the sensitivity of current trypanosome parasitological diagnostic techniques. Bulletin of the World Health Organization 46, 2–7.Google Scholar
Paris, J., Murray, M. & McOdimba, T. (1982). A comparative evaluation of the parasitological techniques currently available for the diagnosis of African trypanosomiasis in cattle. Acta Tropica 39, 307–16.Google ScholarPubMed
Parish, N. M., Morrison, W. I. & Pearson, T. W. (1985). Identification of an antigen specific to Trypanosoma congolense using monoclonal antibodies. Journal of Immunology 134, 593–9.CrossRefGoogle ScholarPubMed
Pearson, T. W., Liu, M. K., Gardiner, I. C., Longridge, D., Beecroft, P. R., Sayer, P. D., Gould, S. S., Waitumbi, J. N. & Njogu, A. R. (1986). Use of procyclic trypanosomes for detection of antibodies in sera from vervet monkeys infected with Trypanosoma rhodesiense: an immunodiagnostic test for African sleeping sickness. Acta Tropica 43, 391–9.Google ScholarPubMed
Peterson, G. L. (1983). Determination of total protein. In Methods in Enzymology. vol. 92, (ed. Hirs, C. H. W. and Timasheff, S. N.), pp. 95–119. New York: Academic Press.Google Scholar
Rae, P. F. & Luckins, A. G. (1984). Detection of circulating trypanosomal antigens by enzyme immunoassay. Annals of Tropical Medicine and Parasitology 78, 587–96.CrossRefGoogle ScholarPubMed
Richardson, J. P., Jenni, L., Beecroft, R. P. & Pearson, T. W. (1986). Procyclic testse fly midgut forms and culture forms of African trypanosomes share stage- and species-specific surface antigens identified by monoclonal antibodies. Journal of Immunology 136, 2259–64.CrossRefGoogle Scholar
Sailyo, R. S., Gray, A. R. & Luckins, A. G. (1980). Use of antigens of cultured Trypanosoma brucei in tests for bovine trypanosomiasis. Tropical Animal Health and Production 12, 127–31.CrossRefGoogle Scholar
Shapiro, S. Z. & Pearson, T. W. (1986). African trypanosomiasis: antigens and host-parasite interactions. In Parasite Antigens: Toward New Strategies for Vaccines (ed. Pearson, T. W.), pp. 215–74. New York and Basel: Marcel Dekker Inc.Google Scholar
Smith, R. D., James, M. A., Ristic, M., Aikawa, M. & Vega, C. A. (1981). Bovine babesiosis: protection of cattle with culture-derived soluble Babesia bovis antigen. Science 212, 335–8.CrossRefGoogle ScholarPubMed
Smith, C. J., Levine, R. F. & Mansfield, J. M. (1982). Cloning of African trypanosomes in mice suppressed by cyclophosphamide treatment. American Journal of Tropical Medicine and Hygiene 31, 1098–102.CrossRefGoogle Scholar
Taylor, A. E. R. & Smith, V. (1983). Micro-counter immunoelectrophoresis: a rapid screening technique for trypanosomiasis. Transactions of the Royal Society of Tropical Medicine and Hygiene 77, 481–6.CrossRefGoogle Scholar
Van Knappen, K., Buys, J. & Ruitenberg, E. J. (1977). Enzyme-linked immunosorbent assay (ELISA) and its application to the serodiagnosis of African trypanosomiasis. Annales de la Société Belge de Médecine Tropicale 57, 281–92.Google Scholar
Van Meirvenne, N. & Le Ray, D. (1985). Diagnosis of African and American trypanosomiases. British Medical Bulletin 42, 156–61.CrossRefGoogle Scholar
Vervoort, T., Magnus, E. & Van Meirvenne, N. (1983). Serological test for sleeping sickness: importance of antigen selection. In Symposium on the Diagnosis of African Sleeping Sickness due to T. gambiense, Anvers-Antwerp, (ed. Crooy, P. J.), pp. 47–50. Belgium: Smith Kline-RIT Biological Division.Google Scholar
Voller, A., Bidwell, D. E. & Bartlett, A. (1975). A serological study of human Trypanosoma rhodesiense infections using a micro-scale enzyme-linked immunosorbent assay. Tropenmedizin und Parasitologie 26, 247–51.Google ScholarPubMed
Wery, M., Wery-Paskoff, S. & Van Wettere, D. (1970). The diagnosis of African trypanosomiasis (T. gambiense) by the use of a fluorescent antibody test. 1. Standardization of an easy technique to be used in mass surveys. Annales de la Société Belge de Médecine Tropicale 50, 613–34.Google Scholar
World Health Organization (1981). Scientific working group on African trypanosomiases. Antwerp.Google Scholar
Wood, J. N. (1984). Subclass analysis and purification of monoclonal antibodies. In Methods in Molecular Biology, vol. 1 (ed. Walker, J. M.), pp. 287–93. New Jersey: The Humana Press.Google Scholar