title: Antigenic components, isolation and partial characterization of excretion-secretion fraction of Paramphistomum cervi.
abstract: The immunogenic components of adult Paramphistomum cervi excretion-secretion (ES) fraction were revealed by SDS-PAGE and immunoblotting technique using sera from cattle naturally infected with P. cervi, Fasciola gigantica, strongylids, Trichuris sp., and Strongyloides sp. By SDS-PAGE, it was found that the ES fraction comprised 13 distinct protein bands. Immunoblotting analysis of these proteins exhibited nine prominent antigenic bands which were recognized by paramphistomosis antisera. These antigenic proteins had molecular weights ranging from 10-170 kDa. One antigenic protein band of 40 kDa was found to give a consistent reaction with sera from all infected cattle. Its diagnostic sensitivity, specificity and accuracy using this test were 100%, 98.9% and 99.3%, respectively. The positive and negative predictive values were 98% and 100%, respectively. The 40 kDa antigen was partially purified by gel filtration and ion-exchange chromatography. The antigenicity of 40 kDa protein for diagnosis of P. cervi infection was confirmed by immunoblotting and indirect ELISA (at 1:78,125 dilution) using a pool of sera and individual serum samples from infected cattle. The present findings suggest that the 40 kDa protein may be used as a diagnostic antigen for paramphistomosis.
authors.affiliation: Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi 71150, Thailand. Panat1@yahoo.com
authors.forename: Panat
authors.initials: P
authors.lastname: Anuracpreeda
authors.forename: Jaruwan
authors.initials: J
authors.lastname: Poljaroen
authors.forename: Charoonroj
authors.initials: C
authors.lastname: Chotwiwatthanakun
authors.forename: Yotsawan
authors.initials: Y
authors.lastname: Tinikul
authors.forename: Prasert
authors.initials: P
authors.lastname: Sobhon
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood / Q000276:immunology; D000947:Antigens, Helminth / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology*; D002850:Chromatography, Gel / Q000662:veterinary; D002852:Chromatography, Ion Exchange / Q000662:veterinary; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D015801:Helminth Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D006374:Helminthiasis, Animal / Q000276:immunology / Q000469:parasitology; D015151:Immunoblotting / Q000662:veterinary; D008970:Molecular Weight; D010250:Paramphistomatidae / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D011237:Predictive Value of Tests; D012417:Rumen / Q000469:parasitology; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000662:veterinary; D014201:Trematode Infections / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins