Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-25T04:34:33.104Z Has data issue: false hasContentIssue false

Enzyme-linked immunosorbent assay, immunoblot analysis and RAST fluoroimmunoassay analysis of serum responses against crude larval antigens of Anisakis simplex in a Spanish random population

Published online by Cambridge University Press:  05 June 2009

L. García-Palacios
Affiliation:
Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain
M.L. González
Affiliation:
Unidad de Alergia, Hospital General de Segovia, Spain
M.I. Esteban
Affiliation:
Unidad de Alergia, Hospital General de Segovia, Spain
E. Mirabent
Affiliation:
Laboratorio de Análisis Clínicos, Hospital General de Segovia, Spain
M.J. Perteguer
Affiliation:
Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain
C. Cuéllar
Affiliation:
Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain

Abstract

The results obtained in a study of seroprevalence by means of ELISA and immunoblot with crude larval extracts of Anisakis simplex using 1008 human sera from Spanish people showing no clinical suspicion of anisakidosis are given. For the evaluation of the results obtained by ELISA the Diagnostic Index (DI) was used, as the ratio between the optical density resulting from the test serum and the optical density of the negative control. Forty-seven sera showed DIs between 1.5 and 2, and 14 sera were greater than 2. After comparison of the immunoblot analysis with the immunorecognition pattern of a human anisakidosis reference serum, a diagnostic criterion could be established for those sera that, at a 1/100 dilution, showed a DI by ELISA greater than 1.5. Seven of 14 selected sera with DIs in ELISA higher than 1.3 showed anti-Anisakis specific IgE antibodies by RAST fluoroimmunoassay.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1996

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

Aguila, C., Cuéllar, C., Fenoy, S. & Guillén, J.L. (1987) Comparative study of assay detecting circulating immuno-complexes and specific antibodies in patients infected with Toxocara canis. journal of Helminthology 61, 196202.CrossRefGoogle Scholar
Amano, T., Nakazawa, M., Sugiyama, H., Secor, W.E. & Oshima, T. (1995) Specific antibody patterns of Wistar rats inoculated with third stage larvae of Anisakis simplex. Journal of Parasitology 81, 536542.Google Scholar
Arenal, J., Marcos, L., Borrego, M., Bowakin, W., Castro, J. & Blanco, J. (1991) Anisakiasis como causa de apendicitis aguda y cuadro reumatológico: primer caso en la literature médica. Revista Española de Enfermedades Digestivas 79, 355358.Google Scholar
Bradford, M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Annals of Biochemistry 72, 248.CrossRefGoogle ScholarPubMed
Clavel, A., Delgado, B., Sanchez-Acedo, C., Carbonell, E., Castillo, J., Ramirez, J., Quilez, J., Gomez, R. & Kagei, N. (1993) A live Anisakis physeteris larva found in the abdominal cavity of a woman in Zaragoza, Spain. Japanese Journal of Parasitology 5, 445448.Google Scholar
Desowitz, R., Raybourne, R., Ishikura, H. & Kliks, M. (1985) The radioallergosorbent test (RAST) for the serological diagnosis of human anisakiasis. Transactions of the Royal Society of Tropical Medicine and Hygiene 79, 256259.CrossRefGoogle ScholarPubMed
Fenoy, S., Cuéllar, C., Aguila, C. & Guillén, J.L. (1989) Posibles reacciones cruzadas del antígeno excretor/secretor de Toxocara canis en ELISA con antígenos de otros ascáridos y proteína C-reactiva. Revista Ibérica de Parasitología 49, 185192.Google Scholar
Hames, B.D. (1986) An introduction to polyacrilamide gel electrophoresis. pp. 2157in Hames, B.D. & Rickwood, D. (Eds) Gel electrophoresis of proteins. Oxford, IRL Press.Google Scholar
Iglesias, R., Leiro, J., Ubeira, F., Santamarina, M. & Sanmartín, M. (1993) Anisakis simplex: antigen recognition and antibody production in experimentally infected mice. Parasite Immunology 15, 243250.CrossRefGoogle ScholarPubMed
Ishikura, H., Kikuchi, K., Nagasawa, K., Ooiwa, T., Takamiya, H., Sato, N. & Sugane, K. (1993) Anisakidae and anisakidosis. pp. 43101in Sun, T. (Ed.) Progress in clinical parasitology. Vol. III. New York, Springer-Verlag.Google Scholar
Kasuya, S., Hamano, H. & Izumi, S. (1990) Mackerel-induced urticaria and Anisakis. Lancet 335, 665.CrossRefGoogle ScholarPubMed
Kennedy, M., Tierney, J., Puzeng, Y., McMonagle, F., McIntosh, A., McLaughlin, D. & Smith, J. (1988) The secreted and somatic antigens of the third stage larva of Anisakis simplex and antigenic relationship with Ascaris suum, Ascaris lumbricoides and Toxocara canis. Molecular and Biochemical Parasitology 31, 3546CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680.CrossRefGoogle ScholarPubMed
Lopez-Velez, R., García, A., Barros, C., Manzabeitia, F. & Oñate, J.M. (1992) Anisakidosis en España. Descripción de tres casos. Enfermedades Infecciosas y Microbiología Clínica 10, 158161.Google Scholar
Perteguer, M.J., Fenoy, S., Cuéllar, C. & Guillén, J.L. (1993) Respuesta inmune humoral en ratones BALB/C inoculados con larvas de tercer estadio de Anisakis. Acta Parasitológica Portuguesa 1, 258.Google Scholar
Petithory, J.C., Verrier, F. & Guyet-Rousset, P. (1992) Biological study of a case of colonic anisakiasis. Review of the literature. Pathologie Biologie 40, 4751.Google Scholar
Sakanari, J.A. & McKerrow, J.H. (1989) Anisakiasis. Clinical Microbiology Reviews 2, 278284.CrossRefGoogle ScholarPubMed
Sakanari, J.A., Loinaz, H.M., Deardorff, T.L., Raybourne, R.B., McKerrow, J.H. & Frieson, J.G. (1988) A case diagnosed by morphological and immunologic methods. American Journal of Clinical Pathology 90, 107113.Google Scholar
Valero, A., Mira, J., Pérez, R., Sanz, J., Geraldía, M., García, J., García, P. & Adroher, F. (1992) Descripción de un caso de anisakiosis humana. IX Reunión Científica de la Asociación de Parasitólogos Españoles, 69.Google Scholar