Purification of larval Taenia solium antigens by isoelectric focusing.
By preparative isoelectric focusing in a rotating ampholine column, crude cystic membrane (M) or fluid (F) antigens of larval Taenia solium were each separated into 20 fractions. M fractions were less specific and sensitive than F fractions in detecting cysticercosis antibodies in pig serum. Among the F fractions, F15 showed the best potential to serve as a screening antigen. It contained 18 polypeptides, with pI 5.3-8.2 and a specific epitope of 25 kDa which was detected by immunoblotting. Although F15 showed slight cross-reactions with heterologous antisera in double-antibody IgG enzyme-linked immunosorbent assays (ELISA), it yielded the highest absorbance values when tested against homologous antisera. The antigen was used to screen sera samples from 4870 pigs slaughtered in Hong Kong and five other Chinese cities for cysticercosis antibodies by double-antibody ELISA, Falcon Assay Screening Test (FAST)-ELISA and enhanced chemiluminescent immunoassay. The results varied significantly between assays. However, the samples collected from Shenzhen yielded the highest positive rates. Enhanced chemiluminescent immunoassay based on camera-luminometry was found suitable for use under field conditions.
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Department of Zoology, University of Hong Kong, China.
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D000665:Ampholyte Mixtures; D000704:Analysis of Variance; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth / Q000097:blood / Q000276:immunology / Q000302:isolation & purification*; D002681:China; D003551:Cysticercosis / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D003552:Cysticercus / Q000737:chemistry / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D006723:Hong Kong; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007525:Isoelectric Focusing / Q000662:veterinary; D008163:Luminescent Measurements; D011817:Rabbits; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000276:immunology* / Q000517:prevention & control; D013621:Taenia / Q000276:immunology*
D000665:Ampholyte Mixtures; D000909:Antibodies, Helminth; D000947:Antigens, Helminth