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Serotype- and serogroup-specific detection of African horsesickness virus using phage displayed chicken scFvs for indirect double antibody sandwich ELISAs
Authors:Wouter van Wyngaardt  Cordelia Mashau  Isabel Wright  Jeanni Fehrsen
Institution:1Immunology Division, Onderstepoort Veterinary Institute, Private Bag X05, Onderstepoort 0110, South Africa.;2Deltamune, P. O. Box 14167, Lyttelton 0140, South Africa.
Abstract:There is an ongoing need for standardized, easily renewable immunoreagents for detecting African horsesickness virus (AHSV). Two phage displayed single-chain variable fragment (scFv) antibodies, selected from a semi-synthetic chicken antibody library, were used to develop double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) to detect AHSV. In the DAS-ELISAs, the scFv previously selected with directly immobilized AHSV-3 functioned as a serotype-specific reagent that recognized only AHSV-3. In contrast, the one selected with AHSV-8 captured by IgG against AHSV-3 recognized all nine AHSV serotypes but not the Bryanston strain of equine encephalosis virus. Serving as evidence for its serogroup-specificity. These two scFvs can help to rapidly confirm the presence of AHSV while additional serotype-specific scFvs may simplify AHSV serotyping.
Keywords:African horsesickness virus  double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA)  phage-displayed scFv  serotype-specific
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