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Fas2-ELISA in the detection of human infection by Fasciola hepatica

Published online by Cambridge University Press:  12 April 2024

J.R. Espinoza*
Affiliation:
Molecular Biotechnology Unit, Laboratories for Research and Development, Faculty of Science and Philosophy, Universidad Peruana Cayetano Heredia, PO Box 4314, Lima, 100, Peru Department of Biology, Peruvian Institute of Nuclear Energy (IPEN), Peru
O. Timoteo
Affiliation:
Molecular Biotechnology Unit, Laboratories for Research and Development, Faculty of Science and Philosophy, Universidad Peruana Cayetano Heredia, PO Box 4314, Lima, 100, Peru
P. Herrera-Velit
Affiliation:
Molecular Biotechnology Unit, Laboratories for Research and Development, Faculty of Science and Philosophy, Universidad Peruana Cayetano Heredia, PO Box 4314, Lima, 100, Peru
*
*Fax: +511 319 0024, E-mail: [email protected]
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Abstract

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Fasciola hepatica has recently emerged as a major pathogen of humans from reports on areas of endemicity and hyper-endemicity for fascioliasis. This situation is aggravated by the lack of standard assays for the screen diagnosis of F. hepatica infection in humans living in endemic areas. Our laboratory has developed an enzyme-linked immunosorbent assay (Fas2-ELISA) based on the capture of IgG antibody by a purified protein Fas2, which is an adult fluke cysteine proteinase. Fas2-ELISA exhibited 95% sensitivity and 100% specificity in 38 individuals infected with F. hepatica diagnosed by finding eggs in stools and 46 serum samples from healthy volunteers. No cross-reaction was observed with 54 serum samples from patients with ten different parasitic infections including the trematodes Paragonimus westermani and Schistosoma mansoni. The high antigenicity of Fas2 is suggested by the fact that antibodies to Fas2 rise rapidly by 1–2 weeks of infection and rise until patency at 8 weeks of infection in experimentally infected alpacas. Field screening for human fascioliasis using Fas2-ELISA and coprology in three endemic locations of the Peruvian Andes resulted in 95.5% sensitivity, 86.6% specificity in a population of 664 children in an age range of 1 to 16 years old. These results provide evidence of the clinical potential of Fas2-ELISA to diagnose fascioliasis in humans exposed to liver fluke infection in endemic areas for this parasite. Fas2-ELISA is currently developed as a standard assay for both field screening for fascioliasis in people living in endemic areas and detecting occasionally F. hepatica infected patients in clinical laboratories.

Type
Review Article
Copyright
Copyright © Cambridge University Press 2005

References

Berasain, P., Goni, F., McGonigle, S., Dowd, A., Dalton, J.P., Frangione, B. & Carmona, C. (1997) Proteinases secreted by Fasciola hepatica degrade extracellular matrix and basement membrane components. Journal of Parasitology 83, 15.CrossRefGoogle ScholarPubMed
Berasain, P., Carmona, C., Frangione, B., Dalton, J.P. & Goni, F. (2000) Fasciola hepatica: parasite-secreted proteinases degrade all human IgG subclasses: determination of the specific cleavage sites and identification of the immunoglobulin fragments produced. Experimental Parasitology 94, 99110.CrossRefGoogle ScholarPubMed
Carmona, C., Dowd, A.J., Smith, A.M. & Dalton, J.P. (1993) Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Molecular and Biochemical Parasitology 62, 917.CrossRefGoogle ScholarPubMed
Carnevale, S., Rodriguez, M.I., Guarnera, E.A., Carmona, C., Tanos, T. & Angel, S.O. (2001) Immunodiagnosis of fasciolosis using recombinant procathepsin L cysteine proteinase. Diagnostic Microbiology and Infectious Disease 41, 4349.CrossRefGoogle Scholar
Chapman, C.B. & Mitchell, G.F. (1982) Proteolytic cleavage of immunoglobulin by enzymes released by Fasciola hepatica . Veterinary Parasitology 11, 165178.CrossRefGoogle ScholarPubMed
Chen, M.G. & Mott, K.E. (1990) Progress in assessment of morbidity due to Fasciola hepatica infection: a review of recent literature. Tropical Diseases Bulletin 87, R1R38.Google Scholar
Cordova, M., Herrera, P., Ñopo, L., Bellatin, J., Náquira, C., Guerra, H. & Espinoza, J.R. (1997) Fasciola hepatica cysteine proteinases: immunodominant antigens in human fascioliasis. American Journal of Tropical Medicine and Hygiene 57, 660666.CrossRefGoogle ScholarPubMed
Cordova, M., Reategui, L. & Espinoza, J.R. (1999) Immunodiagnosis of human fascioliasis with Fasciola hepatica cysteine proteinases. Transactions of the Royal Society of Tropical Medicine and Hygiene 93, 5457.CrossRefGoogle ScholarPubMed
Cornelissen, J.B., Gaasenbeek, C.P., Boersma, W., Borgsteede, F.H. & van Milligen, F.J. (1999) Use of a pre-selected epitope of cathepsin-L1 in a highly specific peptide-based immunoassay for the diagnosis of Fasciola hepatica infections in cattle. International Journal for Parasitology 29, 685696.CrossRefGoogle Scholar
Cornelissen, J.B., Gaasenbeek, C.P., Borgsteede, F.H., Holland, W.G. Harmsen, M.M. & Boersma, W.J. (2001) Early immunodiagnosis of fasciolosis in ruminants using recombinant Fasciola hepatica cathepsin L-like protease. International Journal for Parasitology 31, 728737.CrossRefGoogle ScholarPubMed
Dalton, J.P. & Heffernan, M. (1989) Thiol proteases released in vitro by Fasciola hepatica . Molecular and Biochemical Parasitology 35, 161166.CrossRefGoogle ScholarPubMed
Dalton, J.P., Neill, S.O., Stack, C., Collins, P., Walshe, A., Sekiya, M., Doyle, S., Mulcahy, G., Hoyle, D., Khaznadji, E., Moire, N., Brennan, G., Mousley, A., Kreshchenko, N., Maule, A.G. & Donnelly, S.M. (2003) Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines. International Journal for Parasitology 33, 11731181.CrossRefGoogle ScholarPubMed
Doenhoff, M.J., Chiodini, P.L. & Hamilton, J.W. (2004) Specific and sensitive diagnosis of schistosome infection: can it be done with antibodies?. Trends in Parasitology 20, 3539.CrossRefGoogle ScholarPubMed
Espino, A.M., Marcet, R. & Finlay, C.M. (1990) Detection of circulating excretory secretory antigens in human fascioliasis by sandwich enzyme-linked immunosorbent assay. Journal of Clinical Microbiology 28, 26372640.CrossRefGoogle ScholarPubMed
Espino, A.M., Diaz, A., Perez, A. & Finlay, C.M. (1998) Dynamics of antigenemia and coproantigens during human Fasciola hepatica outbreak. Journal of Clinical Microbiology 36, 27232726.CrossRefGoogle ScholarPubMed
Espino, A.M., Rodriguez-Medina, J.R. & Hillyer, G.V. (2001) Isolation and immunological characterization of fatty acid binding protein isoforms from Fasciola hepatica . Journal of Parasitology 87, 10201033.CrossRefGoogle ScholarPubMed
Halton, D.W. (1967) Observations on the nutrition of digenetic trematodes. Parasitology 57, 639660.CrossRefGoogle ScholarPubMed
Hillyer, G.V., Soler de Galanes, M., Rodríguez-Pérez, J., Bjorland, J., Silva de Lagrava, M., Ramírez Guzmán, S. & Bryan, R.T. (1992) Use of the Falcon assay screening test-enzyme-linked immunosorbent assay (FAST-ELISA) and the enzyme-linked immunoelectrotransfer blot (EITB) to determine the prevalence of human fascioliasis in the Bolivian Altiplano. American Journal of Tropical Medicine and Hygiene 46, 603609.CrossRefGoogle ScholarPubMed
Hillyer, G.V. (1999) Inmunodiagnosis of human and animal fasciolosis. pp. 435447 in Dalton, J.P. (ED.) Fasciolosis . Wallingford, Oxon CABI Publishing.Google Scholar
Kim, K., Yang, J.H. & Chung, Y-B. (2003) Usefu,lness of 8 kDa protein of Fasciola hepatica in diagnosis of fascioliasis. Korean Journal of Parasitology 41, 121123.CrossRefGoogle Scholar
Law, R.H., Smooker, P.M., Irving, J.A., Piedrafita, D., Ponting, R., Kennedy, N.J., Whisstock, J.C., Pike, R.N. & Spithill, T.W. (2003) Cloning and expression of the major secreted cathepsin B-like protein from juvenile Fasciola hepatica and analysis of immunogenicity following liver fluke infection. Infection and Immunity 71, 69216932.CrossRefGoogle ScholarPubMed
Mas-Coma, S., Bargues, M.D. & Esteban, J.G. (1999) Human fasciolosis. pp. 411434 in Dalton, J.P. (ED.) Fasciolosis . allingford, Oxon CABI Publishing.Google Scholar
Mezo, M., Gonzales-Warleta, M., Carro, C. & Ubeira, F.M. (2004) An ultrasensitive capture ELISA for detection of Fasciola hepatica coproantigens in sheep and cattle using a new monoclonal antibody (MM3). Journal of Parasitology 90, 845852.CrossRefGoogle ScholarPubMed
Neyra, V., Chavarry, E. & Espinoza, J.R. (2002) Cysteine proteinases Fas1 and Fas2 are diagnostic markers for Fasciola hepatica infection in alpacas (Lama pacos). Veterinary Parasitology 105, 2132.CrossRefGoogle ScholarPubMed
O'Neill, S.M., Parkinson, M., Strauss, W., Angles, R. & Dalton, J.P. (1998) Immunodiagnosis of Fasciola hepatica infection (fascioliasis) in a human population in the Bolivian Altiplano using purified cathepsin L cysteine proteinase. American Journal of Tropical Medicine and Hygiene 58, 417423.CrossRefGoogle Scholar
O'Neill, S.M., Parkinson, M., Dowd, A.J., Strauss, W., Angles, R. & Dalton, J.P. (1999) Short report: immunodiagnosis of human fascioliasis using recombinant Fasciola hepatica cathepsin L1 cysteine proteinase. American Journal of Tropical Medicine and Hygiene 60, 749751.CrossRefGoogle ScholarPubMed
Paz-Silva, A., Hillyer, G.V., Sánchez-Andrade, R., Rodríguez-Medina, J.R., Arias, M., Morrondo, P. & Diez-Baños, P. (2005) Isolation, identification and expression of a Fasciola hepatica cDNA encoding a 2.9-kDa recombinant protein for the diagnosis of ovine fasciolosis. Parasitology Research 95, 129135.CrossRefGoogle ScholarPubMed
Raymundo, L.A., Flores, V.M., Terashima, A., Samalvides, F., Miranda, E., Tantalean, M., Espinoza, J.R. & Gotuzzo, E. (2004) Hyperendemicity of human fasciolosis in the Mantaro Valley, Peru: factors for infection with Fasciola hepatica . Revista de Gastroenterologia del Peru 24, 158164.Google Scholar
Rokni, M.B., Massoud, J., O'Neill, S.M., Parkinson, M. & Dalton, J.P. (2002) Diagnosis of human fasciolosis in the Gilan province of Northern Iran: application of cathepsin L-ELISA. Diagnostic Microbiology and Infectious Disease 44, 175179.CrossRefGoogle ScholarPubMed
Ruiz, A., Molina, J.M., Gonzales, J., Martinez-Moreno, F.J., Gutierrez, P.N. & Martinez-Moreno, A. (2003) Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke. Veterinary Research 34, 435443.CrossRefGoogle ScholarPubMed
Silva, E., Castro, A., Lopes, A., Rodrigues, A., Dias, C., Conceicao, A., Alonso, J., Correia da Costa, J.M., Bastos, M., Parra, F., Moradas-Ferreira, P. & Silva, M. (2004) A recombinant antigen recognized by Fasciola hepatica -infected hosts. Journal of Parasitology 90, 746751.CrossRefGoogle ScholarPubMed
Smith, A.M., Dowd, A.J., Heffernan, M., Robertson, C.D. & Dalton, J.P. (1993a) Fasciola hepatica: a secreted cathepsin L-like proteinase cleaves host immunoglobulin. International Journal for Parasitology 23, 977983.CrossRefGoogle ScholarPubMed
Smith, A.M., Dowd, A.J., McGonigle, S., Keegan, P.S., Brennan, G., Trudgett, A. & Dalton, J.P. (1993b) Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica . Molecular and Biochemical Parasitology 62, 18.CrossRefGoogle ScholarPubMed
Spithill, T.W., Smooker, P.M., Sexton, J.L., Bozas, E., Morrison, C.A., Creaney, J. & Parsons, J.C. (1999) Development of vaccines against Fasciola hepatica. pp.377410 in Dalton, J.P. (Ed.) Fasciolosis . Wallingford, Oxon CABI Publishing.Google Scholar
Strauss, W., O'Neill, S.M., Parkinson, M., Angles, R. & Dalton, J.P. (1999) Short report: diagnosis of human fascioliasis: detection of anti-cathepsin L antibodies in blood samples collected on filter paper. American Journal of Tropical Medicine and Hygiene 60, 746748.CrossRefGoogle ScholarPubMed
Timoteo, O., Maco, V., Neyra, V., Yi, P.J., Leguia, G. & Espinoza, J.R. (2005) Characterization of the humoral immune response in alpacas (Lama pacos) experimentally infected with Fasciola hepatica against cysteine proteinases Fas1 and Fas2 and histopathological findings. Veterinary Immunology and Immunopathology 106, 7786.CrossRefGoogle ScholarPubMed
Youssef, F.G., Mansour, N.S. & Aziz, A.G. (1991) Early diagnosis of human fascioliasis by the detection of copro-antigens using counterimmunoelectrophoresis. Transactions of the Royal Society of Tropical Medicine and Hygiene 85, 383384.CrossRefGoogle ScholarPubMed