title: Development of a blocking enzyme-linked immunosorbent assay for the detection of avian polyomavirus-specific antibodies.
abstract: Avian polyomavirus, described originally as budgerigar fledgling disease virus, has been associated with devastating contagious disease outbreaks in budgerigar aviaries. At present, this virus affects a wide range of psittacine and non-psittacine birds worldwide, and the serum neutralisation test is used for the serodiagnosis of avian polyomavirus infections. A blocking enzyme-linked immunosorbent assay was developed for the screening of large numbers of sera collected from various avian species. The assay employs a monoclonal antibody directed against the major structural protein VP1 as a blocking antibody in a sandwich blocking procedure. Either purified avian polyomavirus particles or avian polyomavirus VP1 expressed in recombinant baculovirus-infected Sf9 cells were used as antigen. The specificity of the blocking enzyme-linked immunosorbent assay was evaluated by testing sera directed against mammalian polyomaviruses. Using sera obtained from chicken infected experimentally with avian polyomavirus and a collection of psittacine field-origin sera, a good correlation was observed between the results of the blocking enzyme-linked immunosorbent assay and the serum neutralisation test. However, the blocking enzyme-linked immunosorbent assay is more rapid and more economic. Both, avian polyomavirus particles and VP1 produced by recombinant DNA technology proved to be suitable antigens.
authors.affiliation: Institute of Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103, Leipzig, Germany.
authors.forename: M S
authors.initials: MS
authors.lastname: Khan
authors.forename: R
authors.initials: R
authors.lastname: Johne
authors.forename: I
authors.initials: I
authors.lastname: Beck
authors.forename: M
authors.initials: M
authors.lastname: Pawlita
authors.forename: E F
authors.initials: EF
authors.lastname: Kaleta
authors.forename: H
authors.initials: H
authors.lastname: Müller
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D001715:Bird Diseases / Q000175:diagnosis*; D002213:Capsid / Q000276:immunology* / Q000302:isolation & purification; D036022:Capsid Proteins; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007313:Insecta / Q000166:cytology; D009500:Neutralization Tests / Q000379:methods; D011120:Polyomavirus / Q000276:immunology* / Q000302:isolation & purification; D027601:Polyomavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D011563:Psittaciformes; D011994:Recombinant Proteins / Q000276:immunology; D014771:Virion / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D011994:Recombinant Proteins; C062481:VP1 protein, polyomavirus