Parasitology Research

, Volume 78, Issue 2, pp 112–119 | Cite as

Identification of diagnostic antigens fromTrichinella spiralis

  • Wieger L. Homan
  • Anja C. G. Derksen
  • Frans van Knapen
Original Investigations


The Western blotting technique was used to determine the antigens ofTrichinella spiralis muscle larvae that were recognized by antibodies in sera from humans and pigs displayingT. spiralis infections. This resulted in the identification of several antigens that were recognized by all sera. Some of these antigens, notably those that were recognized during the early stage of infection, cross-reacted with antibodies to other parasites. This cross-reactivity was caused by the presence of phosphorylcholine on these antigens. A large portion of the antigens that were recognized by antibodies from infected humans and pigs were found to share a singleTrichinella-specific determinant. TheTrichinella-specific antigen population could be isolated from phosphorylcholine-containing antigens by a simple two-step affinity chromatography procedure using monoclonal antibodies to both determinants. The resulting preparation consisted primarily of a single antigen showing an apparent molecular weight of 45 kDa that corresponded to a mamor constituent of excretory-secretory (ES) products of muscle larvae. When tested in an enzyme-linked immunosorbent assay (ELISA), this antigen displayed diagnostic specificity that was comparable with the ES fraction and diagnostic sensitivity comparable with the crude muscle-larvae extract.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ancelle T, Dupony-Camet J, Heyer F, Faurant C, Lapierre J (1985) Outbreak of trichinosis due to horse meat in the Paris area. Lancet 21:660Google Scholar
  2. Appleton JA, Bell RG, Homan W, Knapen F van (1991) Consensus onTrichinella spiralis Antigens and Antibodies. Parasitol Res 7:190–192Google Scholar
  3. Arriaga C, Muñiz E, Morilla A, Ortega-Pierres G (1989)Trichinella spiralis: recognition of muscle larva antigens during experimental infection of swine and its potential use in diagnosis. Exp Parasitol 69:363–372Google Scholar
  4. Clinard EH (1978) Serum fractions associated with positive and false positive reactions in the EIA test for trichinellosis in swine. In: Kim CW, Pawlowski ZS (eds) Trichinellosis, 4th International Conference, University Press of New England, Hanover, New Hampshire, pp 509–517Google Scholar
  5. Denckers EY, Wassom DL, Hayes CE (1990a) Characterization ofTrichinella spiralis antigens sharing an immunodominant, carbohydrate-associated determinant distinct from phosphorylcholine. Mol Biochem Parasitol 41:241–250Google Scholar
  6. Denckers EY, Wassom DL, Krco CJ, Hayes CE (1990b) The mouse antibody response toTrichinella spiralis defines a single, immunodominant epitope shared by multiple antigens. J Immunol 144:3152–3159Google Scholar
  7. Despommier DD, Laccetti A (1981)Trichinella spiralis: partial characterization of antigens isolated by immuno-affinity chromatography from the large-particle fraction of muscle larvae. J Parasitol 67:332–339Google Scholar
  8. Dupouy-Camet J, Knapen E van, Ancelle T, Vo Quang D, Lavarde V, Lapierre J (1988) Étude des immunoglobulines spécifiques (totales, IgG, IGM, IgA, IgE) en immunofluorescence indirecte et en ELISA chez quarante malades Trichinés suivis pendant neuf mois. Pathol Biol (Paris) 36:803–807Google Scholar
  9. Gamble HR (1985)Trichinella spiralis: immunization of mice using monoclonal antibody affinity-isolated antigens. Exp Parasitol 59:398–404Google Scholar
  10. Gamble HR, Graham CE (1984) Monoclonal antibody-purified antigen for the immunodiagnosis of trichinosis. Am J Vet Res 45:67–74Google Scholar
  11. Gamble HR, Murrell KD (1986) Conservation of diagnostic antigen epitopes among biologically diverse isolates ofTrichinella spiralis. J Parasitol 72:921–925Google Scholar
  12. Gamble HR, Anderson WR, Graham CE, Murrel KD (1983) Diagnosis of swine trichinosis by enzyme-linked immunosorbent assay (ELISA) using an excretory-secretory antigen. Vet Parasitol 13:349–361Google Scholar
  13. Gold AM, Despommier DD, Buck SW (1990) Partial characterization of two antigens secreted by L1 larvae ofTrichinella spiralis. Mol Biochem Parasitol 41:187–196Google Scholar
  14. Knapen F van (1985) The incidence ofTrichinella spiralis in Western Europe in the past decade. Bull Soc Fr Parasitol 2:11–16Google Scholar
  15. Knapen F van (1987) Epidemiological data concerningT. spiralis infections in the Netherlands. Wiad Parazytol 33:566–568Google Scholar
  16. Knapen F van (1988) A propos de cas de trichinose en Europe dus à la consommation de viande chevaline. Ann Med Vet 132:441–446Google Scholar
  17. Knapen F van (1989) Control systems of sylvatic and domestic animals: trichinellosis. Wiad Parazytol 35:475–481Google Scholar
  18. Knapen F van, Franchimont JH, Ruitenberg EJ, Baldelli B, Bradley J, Gibson TE, Gottal C, Henriksen SA, Köhler G, Skovgaard N, Soulé C, Taylor SM (1980) Comparison of the enzyme-linked immunosorbent assay (ELISA) with three methods for the detection ofTrichinella spiralis infections in pigs. Vet Parasitol 7:109–121Google Scholar
  19. Knapen F van, Buys J, Ruitenberg EJ (1986)Trichinella spiralis antibodies. In: Bergmeyer HU, Bergmeyer J, Grassel M (eds) Methods of enzymatic analysis, vol 12. Antigens and antibodies 2. VHC Verlagsgesellschaft, Weinheim, pp 393–406Google Scholar
  20. Kolk AHJ, Ho ML, Klatser PR, Eggelte TA, Kuyper S, Jonge S de, Leetiwen J van (1984) Production and characterisation of monoclonal antibodies toMycobacterium tuberculosis, M. bovis (BCG) andM. leprae. Clin Exp Immunol 58:511–521Google Scholar
  21. Laemmli UK (1970) Cleavage of structural proteins during assembly of bacteriophage T4. Nature 227:680–685Google Scholar
  22. Lal RB, Ottesen EA (1989) Phosphorylcholine epitopes on helminth and protozoal parasites and their presence in the circulation of infected human patients. Trans R Soc Trop Med Hyg 83:622–655Google Scholar
  23. McLaren DJ, Ortega-Pierres G, Parkhouse RME (1987)Trichinella spiralis: immunocytochemical localisation of surface and intracellular antigens using monoclonal antibody probes. Parasitology 94:101–104Google Scholar
  24. Morrissey JH (1981) Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310Google Scholar
  25. O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021Google Scholar
  26. Ortega-Pierres G, Muñiz E, Coral-Vázquez R, Parkhouse RME (1989) Protection againstTrichinella spiralis induced by purified stage-specific surface antigens of infective larvae. Parasitol Res 75:563–567Google Scholar
  27. Pawlowski Z, Knapen F van, Kociecka W, Stefaniak J, Franchimont JH (1990) Clinical expression of trichinellosis and immunoglobulins in three different epidemics. Bull Soc Fr Parasitol 8:996Google Scholar
  28. Perry P, Petit A, Poulain J (1974) Phosphorylcholine-bearing components in homogenates of nematodes. Eur J Immunol 4:637–639Google Scholar
  29. Roach TIA, Wakelin D, Else KJ, Bundy DAP (1988) Antigenic cross-reactivity between the human whipwormTrichuris trichiura and the mouse trichuroidsTrichuris muris andTrichinella spiralis. Parasite Immunol 10:279–291Google Scholar
  30. Ruitenberg EJ, Knapen J van (1977) Enzyme-linked immunosorbent assay (ELISA) as a diagnostic method forTrichinella spiralis infections in pigs. Vet Parasitol 3:317–326Google Scholar
  31. Ruitenberg EJ, Knapen F van (1983) Report 1987–1982 concerningTrichinella spiralis studies in the Netherlands. Wiad Parazytol 29:624–626Google Scholar
  32. Ruitenberg EJ, Lungström I, Steerenberg PA, Buys J (1976) Reliability of the enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis ofTrichinella spiralis infections in conventially raised pigs. J Immunol Methods 10:57–70Google Scholar
  33. Seawright GL, Despommier D, Zimmerman W, Isenstein RS (1983) Enzyme immunoassay for swine trichinelosis using antigens purified by immunoaffinity chromatography. Am J Trop Med Hyg 32:1275–1284Google Scholar
  34. Silberstein DS, Despommier DD (1984) Antigens fromTrichinella spiralis that induce a protective response in the mouse. J Immunol 132:898–904Google Scholar
  35. Taylor SM, Kenny T, Mallon T, Davidson WB (1980) The micro-ELISA for antibodies toTrichinella spiralis: elimination of false positive reactions by antigen fractionation and technical improvements. Z Veterinaermed 27:764–772Google Scholar
  36. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354Google Scholar
  37. Ubeira FM, Leiro J, Santamarina MT, Villa TG, Sanmartín-Durán ML (1987) Immune response toTrichinella epitopes: the antiphosphorylcholine plaque-forming cell response during the biological cycle. Parasitology 94:543–553Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Wieger L. Homan
    • 1
  • Anja C. G. Derksen
    • 1
  • Frans van Knapen
    • 1
  1. 1.Laboratory of Parasitology and MycologyNational Institute of Public Health and Environmental ProtectionBilthovenThe Netherlands

Personalised recommendations