Serologic recognition of hydatid cyst antigens using different purification methods.
The specificity and sensitivity of enzyme-linked immunosorbent assays (ELISA) and Western immunoblot assays in detecting antibodies in serum from patients suffering cystic hydatid disease (Echinococcus granulosus) are compared using either crude antigen preparations (total sheep hydatid fluid and homogenates of protoscoleces), purified fractions enriched in Antigens 5 and B, and glycoproteins from hydatid fluid. Polyprotein bands of 12-14, 20, and 34 kDa, when purified from hydatid fluid by applying changes in the ionic strength, yielded a sensitive (95%) immunodiagnostic test that was also extremely specific (100%) when assayed with sera from noninfected humans and from patients suffering from other parasitic diseases. However, subjecting hydatid fluid to chromatography through a concanavilin A column rendered a 42 kDa band that was sensitive (95%) as well as highly specific (100%) for hydatidosis. Therefore purification procedures can strongly affect the diagnostic value of antigens with identical electrophoretic behavior in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
Sbihi
Y
Y
Instituto de Biotecnología, Universidad de Granada, Spain.
Janssen
D
D
Osuna
A
A
D000818:Animals; D000909:Antibodies, Helminth / Q000032:analysis*; D000947:Antigens, Helminth / Q000302:isolation & purification*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003551:Cysticercosis / Q000276:immunology; D003560:Cysts / Q000276:immunology; D004444:Echinococcosis, Hepatic / Q000175:diagnosis* / Q000276:immunology*; D004446:Echinococcus / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D007896:Leishmaniasis / Q000276:immunology; D012680:Sensitivity and Specificity; D012756:Sheep; D014020:Tissue Extracts / Q000276:immunology; D014123:Toxoplasmosis / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D006023:Glycoproteins; D014020:Tissue Extracts