Characterization and optimization of bovine Echinococcus granulosus cyst fluid to be used in immunodiagnosis of hydatid disease by ELISA.

The aim of this work was to assess the influence in the diagnostic value for human hydatid disease of the composition of bovine hydatid cyst fluid (BHCF) obtained from fertile (FC) and non-fertile cysts (NFC). Eight batches from FC and 5 from NFC were prepared and analysed with respect to chemical composition: total protein, host-derived protein, carbohydrate and lipid contents. No differences were observed in the first two parameters but carbohydrate and lipid contents were shown to be higher in batches from FC than in those from NFC. Bands of 38 and 116 kD in SDS-PAGE profiles were observed to be present in BHCF from FC only. Two pools were prepared from BHCF batches obtained from FC (PFC) and NFC (PNFC), respectively. Antigen recognition patterns were analysed by immunoblot. Physicochemical conditions for adsorption of antigens to the polystyrene surface (ELISA plates) were optimized. The diagnostic value of both types of BHCF as well as the diagnostic relevance of oxidation of their carbohydrate moieties with periodate were assessed by ELISA using 42 serum samples from hydatid patients, 41 from patients with other disorders, and 15 from healthy donors. Reactivity of all sera against native antigen were tested with and without free phosphorylcholine. The best diagnostic efficiency was observed using BHCF from periodate-treated PFC using glycine buffer with strong ionic strength to coat ELISA plates.

[1]  A. Nieto,et al.  Diagnostic evaluation of a synthetic peptide derived from a novel antigen B subunit as related to other available peptides and native antigens used for serology of cystic hydatidosis , 1998, Parasite immunology.

[2]  A. Nieto,et al.  Modified ELISA for hydatid serodiagnosis: the potential of periodate treatment and phosphorylcholine inhibition , 1997 .

[3]  R. Riganò,et al.  Immunological responses to antigen B from Echinococcus granulosus cyst fluid in hydatid patients , 1996, Parasite immunology.

[4]  M. Miguez,et al.  Carbohydrates on the surface of Echinococcus granulosus protoscoleces are immunodominant in mice , 1996, Parasite immunology.

[5]  A. Nieto,et al.  Antibody response of Echinococcus granulosus infected mice: recognition of glucidic and peptidic epitopes and lack of avidity maturation , 1996, Parasite immunology.

[6]  A. Nieto,et al.  High performance latex reagent for hydatid serology using an Echinococcus granulosus lipoprotein antigen fraction purified from cyst fluid in one step. , 1993, International journal for parasitology.

[7]  K. Behbehani,et al.  Specificity of counterimmunoelectrophoresis with an arc-5 antigen for the diagnosis of cystic hydatid disease , 1993 .

[8]  M. Kamboj,et al.  Specific antibodies in serum of patients with hydatidosis recognised by immunoblotting. , 1992, Journal of medical microbiology.

[9]  A. Nieto,et al.  Metalloproteinases in the larvae of Echinococcus granulosus. , 1991, International journal for parasitology.

[10]  R. Riganò,et al.  Detection of antibodies against Echinococcus granulosus major antigens and their subunits by immunoblotting. , 1991, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[11]  G. Bahr,et al.  Diagnostic value of anti-arc 5 IgG antibody and analysis of the IgG subclasses in sera of patients with cystic hydatid disease. , 1990 .

[12]  M. Lightowlers,et al.  Subunit composition and specificity of the major cyst fluid antigens of Echinococcus granulosus. , 1989, Molecular and biochemical parasitology.

[13]  H. Carol,et al.  Lack of interspecies barriers in anti–Id stimulated antibody production against Echinococcus granulosus antigens , 1989, Parasite immunology.

[14]  A. Grabowska,et al.  Human antibody responses to Schistosoma mansoni: the influence of epitopes shared between different life‐cycle stages on the response to the schistosomulum , 1988, European journal of immunology.

[15]  S. Pauluzzi,et al.  The Behaviour of Specific Antibody Classes in Human Hydatid Disease , 1988, Pathology.

[16]  D. McManus,et al.  Specific and cross-reactive antigens of Echinococcus granulosus hydatid cyst fluid. , 1987, Molecular and biochemical parasitology.

[17]  M. Piens,et al.  Host serum protein levels in cysts of human hydatidosis. , 1987, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[18]  A. Nieto,et al.  New experimental criteria for optimization of solid-phase antigen concentration and stability in ELISA. , 1986, Journal of immunological methods.

[19]  J. D. Smyth,et al.  Hydatidosis: changing concepts in epidemiology and speciation. , 1986, Parasitology today.

[20]  Cambridge Ma,et al.  Hydatidosis: Changing Concepts in Epidemiology and, Speciation , 1986 .

[21]  J. Kennedy,et al.  Carbohydrate analysis: a practical approach , 1986 .

[22]  J. D. Smyth,et al.  The Biology of Echinococcus and hydatid disease , 1986 .

[23]  Robert A. Bloodgood,et al.  Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. , 1985, Journal of immunological methods.

[24]  C. Arme,et al.  Echinococcus granulosus: failure of the eosin-exclusion test to demonstrate death of protoscoleces. , 1985, Annals of tropical medicine and parasitology.

[25]  A. Nieto,et al.  Direct measurement of antibody affinity distribution by hapten-inhibition enzyme immunoassay. , 1984, Molecular immunology.

[26]  M. Rickard.  SEROLOGICAL DIAGNOSIS AND POST‐OPERATIVE SURVEILLANCE OF HUMAN HYDATID DISEASE. I. LATEX AGGLUTINATION AND IMMUNOELECTROPHORESIS USING CRUDE CYST FLUID ANTIGEN , 1984, Pathology.

[27]  V. Tsang,et al.  Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. , 1983, Methods in enzymology.

[28]  M. Dovis,et al.  ELISA for antibody measurement: aspects related to data expression. , 1982, Journal of immunological methods.

[29]  A. Farr,et al.  Immunohistochemistry with enzyme labeled antibodies: a brief review. , 1981, Journal of immunological methods.

[30]  J. Guisantes,et al.  Comparison of three immunological tests for seroepidemiological purposes in human echinococcosis , 1981, Parasite immunology.

[31]  H. Lenhoff,et al.  A sensitive and versatile chromogenic assay for peroxidase and peroxidase-coupled reactions. , 1980, Analytical biochemistry.

[32]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[33]  A Voller,et al.  Enzyme immunoassays in diagnostic medicine. Theory and practice. , 1976, Bulletin of the World Health Organization.

[34]  L. Yarzábal,et al.  Evaluation of four variants of the indirect hemagglutination test for human hydatidosis. , 1975, The American journal of tropical medicine and hygiene.

[35]  L. Yarzábal,et al.  Evaluation of whole and purified hydatid fluid antigens in the diagnosis of human hydatidosis by the immunoelectrophoresis test. , 1975, The American journal of tropical medicine and hygiene.

[36]  D. Ridley,et al.  Immunodiagnosis of parasitic diseases. , 1971, The Practitioner.

[37]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[38]  O. Panamericana.  Centro Panamericano de Zoonosis , 1957 .