Parasitology Research

, Volume 77, Issue 8, pp 697–702 | Cite as

Application of the Western blotting procedure for the immunodiagnosis of human toxocariasis

  • J. -F. Magnaval
  • R. Fabre
  • P. Maurières
  • J. -P. Charlet
  • B. de Larrard
Original Investigations


To improve the immunodiagnosis of human toxocaral disease, a sensitive and specific assay using the Western blotting procedure (WB) with excretory-secretory antigens fromToxocara canis larvae (TES) was developed and compared with the standard enzymelinked immunosorbent assay method (TES-ELISA) using the same antigens. We tested groups of sera from laboratory animals or patients presenting with toxocariasis or other helminthic diseases and a group of sera from people dwelling in an area endemic for toxocariasis who exhibited hypereosinophilia. Statistically, the WB assay correlated well with TES-ELISA, but the former was more specific for banding patterns corresponding to low-molecular-weight fractions, thus avoiding problems of cross-reactivity with sera infected with other helminthic diseases.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Akao N, Kondo K, Okamoto T, Yoshimura H (1983) Antigenic analysis of exceretory-secretory products of 2nd stage larvae ofToxocara canis and the antigen recognition in the course of infection. Jpn J Parasitol 32:541–548Google Scholar
  2. Badley JE, Grieve RB, Bowman DD, Glickman LT, Rockey JH (1987) Analysis ofToxocara canis larval excretory-secretory antigens: physico-chemical characterization and antibody recognition. J Parasitol 73:593–600Google Scholar
  3. Beaver PC, Snyder CH, Carrera GM, Dent JH, Lafferty JW (1952) Chronic eosinophilia due to visceral larva migrans. Pediatrics 9:7–19Google Scholar
  4. Bowman DD, Mika-Grieve M, Grieve RB (1987) Circulating excretory-secretory antigen levels and specific antibody responses in mice infected withToxocara canis. Am J Trop Med Hyg 36:75–82Google Scholar
  5. Burnette WN (1981) “Western-blotting”: electrophoretic transfer of proteins from sodium dodecyl sulphate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203Google Scholar
  6. Carlier Y, Yang J, Bout D, Capron A (1982) The use of an excretory-secretory antigen for an ELISA-specific serodiagnosis of visceral larva migrans. Biomed Pharmacother 36:39–42Google Scholar
  7. De Savigny DH (1975) In vitro maintenance ofToxocara canis larvae and a simple method for the production ofToxocara ES antigens for use in serodiagnostic tests for visceral larva migrans. J Parasitol 61:781–782Google Scholar
  8. De Savigny DH, Voller A, Woodruff AW (1979) Toxocariasis: serological diagnosis by enzyme immuno-assay. J Clin Pathol 32:284–288Google Scholar
  9. Diday E (1971) La méthode des nuées dynamiques. Rev Stat Appl 19:19–34Google Scholar
  10. Esteban JL, Chang Chin T, Kay JWD (1985) Importance of Western blot analysis in predicting infectivity of anti-HTLV III/LAV positive blood. Lancet II:1083–1086Google Scholar
  11. Glickman LT, Schantz PM (1985) DoToxocara canis larval antigens used in enzyme-linked immunosorbent assay for visceral larva migrans cross-react with AB isohemagglutinins and give false positive results? Z Parasitenkd 71:395–400Google Scholar
  12. Glickman LT, Schantz PM, Dombroske ER, Cypess R (1978) Evaluation of serodiagnostic tests for visceral larva migrans. Am J Trop Med Hyg 27:492–498Google Scholar
  13. Glickman LT, Magnaval, J-F, Domanski LM, Shofer FS, Lauria SS, Gottstein B, Brochier B (1987) Visceral larva migrans in French adults: a new disease syndrome? Am J Epidemiol 125:1019–1034Google Scholar
  14. Gottstein B, Schantz PM, Todorov T, Saimat AG, Jacquier P (1986) An international study on the serological differential diagnosis of human cytic and alveolar echinococcosis. Bull WHO 64:101–105Google Scholar
  15. Kennedy MW, Maizels RM, Meghji M, Young L, Quereshi F, Smith HV (1987) Species-specific and common epitopes on the secreted and surface antigens ofT. canis andT. cati infective larvae. Parasite Immunol 70:522–529Google Scholar
  16. Lowry O, Rosenbrough NT, Farr A, Randall A (1951) Protein measurement with Folin reagent. J Biol Chem 193:265–275Google Scholar
  17. Lynch NR, Wilkes LK, Hodgen AN, Turner KJ (1988) Specificity ofToxocara ELISA in tropical populations. Parasite Immunol 10:323–337Google Scholar
  18. Magnaval J-F (1987) Eléments nouveaux dans la séméiologie des larva migrans viscérales. Presse Med 16:151–154Google Scholar
  19. Magnaval J-F, Brochier B, Charlet J-P, Larrouy G, Gonzaga dos Santos L (1985) Depistage de la maladie de Chagas par immunoenzymologie. Comparaison de l'ELISA avec l'immunofluorescence et l'hémagglutination indirecte chez 976 donneurs de sang. Rev Transfus Immuno-Hematol 20:201–213Google Scholar
  20. Maizels RM, De Savigny DH, Oglivie BM (1984) Characterization of surface and excretory-secretory antigens ofToxocara canis infective larvae. Parasite Immunol 6:23–27Google Scholar
  21. Nicholas WL, Stewart AC, Mitchell GF (1984) Antibody response toToxocara canis using sera from parasite infected mice and protection from toxocariasis by immunisation with ES antigens. Austr J Exp Biol Med Sci 62:619–626Google Scholar
  22. Nichols RL (1956) The etiology of visceral larva migrans. Diagnostic morphology of infective 2nd-stageToxocara larvae. J Parasitol 42:349–362Google Scholar
  23. Schantz PM (1989)Toxocara larva migrans now. Am J Trop Med Hyg 41 [Suppl]:21–34Google Scholar
  24. Speiser F, Gottstein B (1984) A collaborative study on larval secretory-excretory antigens ofToxocara canis for the immunodiagnosis of human toxocariasis with ELISA. Acta Trop (Basel) 41:361–372Google Scholar
  25. Sugane K, Oshima T (1983) Purification and characterization of excretory-secretory antigens ofToxocara canis larvae. Immunology 50:113-;20Google Scholar
  26. Taylor MR, Keane CT, O'Connor P, Mulvihill E, Holland C (1988) The expanded spectrum of toxocaral disease. Lancet I:692–694Google Scholar
  27. Tsang VCW, Peralta JM, Simmons AR (1983) Enzyme-linked immunoelectrotransfer blot techniques for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods Enzymol 92:377–391Google Scholar
  28. Van Knapen F, Van Leusden J, Polderman AM, Franchimont JH (1983) Visceral larva migrans: examination by means of enzyme-linked immunosorbent assay of human sera for antibodies to excretory-secretory antigens of the second-stage larvae ofToxocara canis. Z Parasitenkd 69:113–118Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • J. -F. Magnaval
    • 1
  • R. Fabre
    • 1
  • P. Maurières
    • 1
  • J. -P. Charlet
    • 1
  • B. de Larrard
    • 1
  1. 1.Laboratoire de ParasitologieCHU Toulouse PurpanToulouseFrance

Personalised recommendations