Hostname: page-component-84b7d79bbc-c654p Total loading time: 0 Render date: 2024-07-27T06:17:45.177Z Has data issue: false hasContentIssue false

Evaluation of purified 27.5 kDa protoscolex antigen-based ELISA for the detection of circulating antigens and antibodies in sheep and human hydatidosis

Published online by Cambridge University Press:  09 July 2014

I.R. Bauomi
Affiliation:
Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt
A.M. El-Amir
Affiliation:
Immunology Department, Faculty of Science, Cairo University, Egypt
A.M. Fahmy
Affiliation:
Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt
R.S. Zalat
Affiliation:
Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt
T.M. Diab*
Affiliation:
Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt

Abstract

Hydatidosis is a zoonotic disease caused by the larval stage of Echinococcus granulosus, and the diagnosis of hydatidosis to date remains unresolved despite the development of many serological techniques. The present study aimed to develop an antigen-based enzyme-linked immunosorbent assay (ELISA) using IgG anti-27.5 kDa protoscolex antigen (27.5 PA) for measuring circulating protoscolex antigen (CPA), for comparison with an antibody detection assay, in sera of naturally infected sheep and humans in highly endemic areas in Egypt. In sheep, the sensitivity of ELISA in detecting anti-27.5 PA IgG and CPA was 75.0 and 60.0%, respectively, and the recorded specificity was 80.0 and 88.0%, respectively. In humans, the sensitivity of ELISA in detecting anti-27.5 PA IgG and CPA was 62.5 and 52.5%, respectively, while the specificity of the assay was 66.7 and 75.0%, respectively. In conclusion, an antibody detection assay is still superior and is more sensitive than an antigen detection assay, especially in diagnosing an active infection in which hydatid cysts are predominant. An antigen detection assay may be a useful approach to assessment of the efficacy of treatment, especially after removal of the cyst. Further studies are recommended to improve the diagnostic efficacy of an antigen-based ELISA method by using a highly purified recombinant antigen.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2014 

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

Allam, G., Bauomy, I.R., Hemyeda, Z.M., Diab, T.M. & Sakran, T.F. (2012) Diagnostic potential of Fasciola gigantica-derived 14.5 kDa fatty acid binding protein in the immunodiagnosis of bubaline fascioliasis. Journal of Helminthology 87, 147153.CrossRefGoogle ScholarPubMed
Beggs, I. (1985) The radiology of hydatid disease. American Journal of Roentgenology 145, 639648.CrossRefGoogle ScholarPubMed
Ben Nouir, N., Gianinazzi, C., Gorcii, M., Müller, N., Nouri, A.L., Babbaa, H. & Gottstein, B. (2009) Isolation and molecular characterization of recombinant P29 protein (recP29) and its assessment for the post-surgical serological follow-up of human cystic echinococcosis in young patients. Transactions of the Royal Society of Tropical Medicine and Hygiene 103, 355364.CrossRefGoogle ScholarPubMed
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.CrossRefGoogle ScholarPubMed
Brunicardi, F.C. (2005) Liver. pp. 11391187in Brunicardi, F.C., Andersen, D.K. & Billiar, T.R. (Eds) Schwartz's principles of surgery. 8th edn.Philadelphia, McGraw-Hill.Google Scholar
Budke, C.M., Deplazes, P. & Torgerson, P.R. (2006) Global socioeconomic impact of cystic echinococcosis. Emerging Infectious Diseases 12, 296303.CrossRefGoogle ScholarPubMed
Carmena, D., Benito, A. & Eraso, E. (2006) Antigens for the immunodiagnosis of Echinococcus granulosus infection: an update. Acta Tropica 98, 7486.CrossRefGoogle ScholarPubMed
Chemale, G., Van Rossum, A.J., Jefferies, J.R., Barrett, J., Brophy, P.M. & Ferreira, H.B. (2003) Proteomic analysis of the larval stage of the parasite Echinococcus granulosus: causative agent of cystic hydatid disease. Proteomics 3, 16331636.CrossRefGoogle ScholarPubMed
Craig, P.S. & Nelson, G.S. (1984) The detection of circulating antigen in human hydatid disease. Annals of Tropical Medicine and Parasitology 78, 219227.CrossRefGoogle ScholarPubMed
Devi, C.S. & Parija, S.C. (2003) A new serum hydatid antigen detection test for diagnosis of cystic echinococcosis. American Journal of Tropical Medicine and Hygiene 69, 525528.CrossRefGoogle ScholarPubMed
Doiz, O., Benito, R., Sbihi, Y., Osuna, A., Clavel, A. & Gomez-Lus, R. (2001) Western blot applied to the diagnosis and post-treatment monitoring of human hydatidosis. Diagnostic Microbiology and Infectious Disease 41, 139142.CrossRefGoogle Scholar
Eckert, J. & Deplazes, P. (2004) Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clinical Microbiology Reviews 17, 107135.CrossRefGoogle ScholarPubMed
Eckert, J. & Thompson, R.C.A. (1997) Intraspecific variation of Echinococcus granulosus and related species with emphasis on their infectivity to humans. Acta Tropica 64, 1934.CrossRefGoogle ScholarPubMed
Engvall, E. & Perlman, P. (1971) Enzyme linked immunosorbent assay (EL1SA). Quantitative assay of immunuglobulin G. Journal of Immunoassay and Immunochemistry 8, 871874.CrossRefGoogle Scholar
Fagbemi, B.O., Obarisiagbon, I.O. & Mbuh, J.V. (1995) Detection of circulating antigen in sera of Fasciola gigantica-infected cattle with antibodies reactive with a Fasciola-specific 88-kDa antigen. Veterinary Parasitology 58, 235246.CrossRefGoogle ScholarPubMed
Galen, R.S. (1980) Predictive values and efficiency of laboratory testing. Pediatric Clinics of North America 27, 861869.CrossRefGoogle ScholarPubMed
Gasser, R.B., Lightowlers, M.W. & Rickard, M.D. (1989) Identification of protein components of Echinococcus granulosus protoscolex antigens for specific serodiagnosis of Echinococcus granulosus in dogs. Parasite Immunology 11, 279291.CrossRefGoogle ScholarPubMed
Gasser, R.B., Parada, L., Acuna, A., Burges, C., Laurenson, M.K., Gulland, F.M., Reichel, M.P. & Paolillo, E. (1994) Immunological assessment of exposure to Echinococcus granulosus in a rural dog population in Uruguay. Acta Tropica 58, 179185.CrossRefGoogle Scholar
González, G., Spinelli, P., Lorenzo, C., Hellman, U., Nieto, A., Willis, A. & Salinas, G. (2000) Molecular characterization of P-29, a metacestode-specific component of Echinococcus granulosus which is immunologically related to, but distinct from, antigen 5. Molecular and Biochemical Parasitology 105, 177184.CrossRefGoogle ScholarPubMed
Gottstein, B. (1984) An immunoassay for the detection of circulating antigens in human echinococcosis. American Journal of Tropical Medicine and Hygiene 33, 11851191.CrossRefGoogle ScholarPubMed
Guohadia, E.E. & Fagbemi, B.O. (1997) The isolation of Fasciola gigantica-specific antigens their use in the serodiagnosis of fasciolosis in sheep by the detection of circulating antigens. Veterinary Parasitology 68, 269282.Google Scholar
Khuroo, M.S. (2002) Hydatid disease: current status and recent advances. Annals of Saudi Medicine 22, 12.CrossRefGoogle ScholarPubMed
Kittelberger, R., Reichel, M.P., Jenner, J., Heath, D.D., Lightowlers, M.W., Moroe, P., Ibrahem, M.M., Craig, P.S. & O'Keefe, J.S. (2002) Evaluation of three enzyme-linked immunosorbent assays (ELISAs) for the detection of serum antibodies in sheep infected with Echinococcus granulosus. Veterinary Parasitology 110, 5776.CrossRefGoogle Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277, 680685.CrossRefGoogle Scholar
Lymbery, A.J. & Thompson, R.C. (1996) Species of Echinococcus: pattern and process. Parasitology Today 12, 486491.CrossRefGoogle ScholarPubMed
McKinney, M.M. & Parkinson, A. (1987) A simple, non-chromatographic procedure to purify immunoglobulins from serum and ascites fluid. Journal of Immunological Methods 96, 271278.CrossRefGoogle ScholarPubMed
Moro, P. & Schantz, P.M. (2009) Echinococcosis: a review. International Journal of Infectious Disease 13, 125133.CrossRefGoogle ScholarPubMed
Nowotny, A. (1979) Basic exercises in immunochemistry. pp. 720. NewYork, Springer Verlag.CrossRefGoogle Scholar
Pawlowski, Z.S., Eckert, J., Vuitton, D.A., Ammann, R.W., Kern, P. & Craig, P.S. (2001) Echinococcosis in humans: clinical aspects, diagnosis and treatment. pp. 2071in Eckert, J., Gemmell, M.A., Meslin, F.X. & Pawlowski, Z.S. (Eds) WHO/OIE manual on echinococcosis in humans and animals: a public health problem of global concern. Paris, WHO/OIE.Google Scholar
Poretti, D., Felleisen, E., Grimm, F., Pfister, M., Teuscher, F., Zuercher, C., Reichen, J. & Gottstein, B. (1999) Differential immunodiagnosis between cystic hydatid disease and other cross-reactive pathologies. American Journal of Tropical Medicine and Hygiene 60, 193198.CrossRefGoogle ScholarPubMed
Rafiei, A. & Craig, P.S. (2002) The immunodiagnostic potential of protoscolex antigens in human cystic echinococcosis and the possible influence of parasite strain. Annals of Tropical Medicine and Parasitology 96, 383389.CrossRefGoogle ScholarPubMed
Ravinder, P.T., Parija, S.C. & Rao, K.S. (1997) Evaluation of human hydatid disease before and after surgery and chemotherapy by demonstration of hydatid antigens and antibodies in serum. Journal of Medical Microbiology 47, 859864.CrossRefGoogle Scholar
Ris, D.R., Hamel, K.L. & Mackle, Z.M. (1987) Use of two polysaccharide antigens in ELISA for the detection of antibodies to Echinococcus granulosus in sheep sera. Research in Veterinary Science 43, 257263.CrossRefGoogle Scholar
Sadjjadi, S.M., Abidi, H., Sarkari, B., Izadpanah, A. & Kazemian, S. (2007) Evaluation of enzyme-linked immunosorbent assay, utilizing native antigen B for serodiagnosis of human hydatidosis. Iranian Journal of Immunology 4, 167172.Google ScholarPubMed
Sadjjadi, S.M., Sedaghat, F., Hosseini, S.V. & Sarkari, B. (2009) Serum antigen and antibody detection in echinococcosis: application in serodiagnosis of human hydatidosis. Korean Journal of Parasitology 47, 153157.CrossRefGoogle ScholarPubMed
Sapero, J.J. & Lawless, D.K. (1953) The MIF stain preservative technique for the identification of intestinal parasites. American Journal of Tropical Medicine and Hygiene 2, 613619.CrossRefGoogle Scholar
Sarkari, B., Sadjjadi, S.M., Abidi, H., Izadpanah, A., Kazemian, S. & Rafati, A. (2007) Application of western blotting using native antigen B for serodiagnosis of human cystic echinococcosis. Iranian Journal of Parasitology 2, 712.Google Scholar
Shariff, M. & Parija, S.C. (1993) Co-agglutination (Co-A) test for circulating antigen in hydatid disease. Journal of Medical Microbiology 38, 391394.CrossRefGoogle ScholarPubMed
Siavashi, M.R., Taherkhani, H., Rezaei, K.R., Razavi Deligani, M.R. & Assmar, M. (2005) Comparison of dot-ELISA and sandwich ELISA diagnostic tests in detection of human hydatidosis. Iranian Biomedical Journal 9, 9194.Google Scholar
Siongok, T.K.A., Mahmoud, A.A.F., Guma, J.H., Warren, K.S., Muller, A.S., Handa, A.K. & Houser, H.B. (1976) Morbidity in schistosomiasis mansoni in relation to intensity of infection: a study of a community in Machakos, Kenya. American Journal of Tropical Medicine and Hygiene 25, 273284.CrossRefGoogle Scholar
Sunita, T., Dubey, M.L., Khurana, S. & Malla, N. (2007) Specific antibody detection in serum, urine and saliva samples for the diagnosis of cystic echinococcosis. Acta Tropica 101, 187191.CrossRefGoogle ScholarPubMed
Tijssen, P. & Kurstak, P. (1984) Highly efficient and simple methods for the preparation of peroxidase and active peroxidase–antibody conjugate for enzyme immunoassays. Analytical Biochemistry 136, 451457.CrossRefGoogle ScholarPubMed
Torgerson, P.R. & Deplazes, P. (2009) Echinococcosis: diagnosis and diagnostic interpretation in population studies. Trends in Parasitology 25, 164170.CrossRefGoogle ScholarPubMed
Woodward, M., Young, W. & Bloodgood, R. (1985) Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. Journal of Immunological Methods 78, 143153.CrossRefGoogle ScholarPubMed
Zhang, W., Li, J. & McManus, D.P. (2003) Concepts in immunology and diagnosis of hydatid disease. Clinical Microbiology Reviews 16, 1836.CrossRefGoogle ScholarPubMed