title: Evaluation of a standard immunofluorescence assay and a new flow cytometric method for the detection of autoreactive antibodies in dogs with tumours.
abstract: The major purpose of the presented study was to develop and to evaluate a flow cytometry-based assay (IIFC) for the determination of autoreactive antibodies in sera from canine cancer patients. A blinded study demonstrated the poor reproducibility of the standard, slide-based and microscopically evaluated indirect immunofluorescence test (IIF), especially with sera displaying a cytoplasmic reactivity. In the IIFC, the intra assay coefficient of variance ranged between 5% and 11%, the inter assay variance between 8% and 25%. The IIFC resulted in significantly less positive results among canine cancer patients (16%) than the IIF (40%). The latter results were due to low titered sera indicating that the standard assay may lead to a high proportion of false positive results. The limitation of the IIFC is that no conclusions can be made about the sub cellular localization of the fluorescence. However, this cytometry-based assay makes a more objective and standardized detection of canine autoreactive antibodies possible.
authors.affiliation: Institute for Immunology, University of Veterinary Medicine, D-30173 Hannover, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Rösner
authors.forename: R
authors.initials: R
authors.lastname: Mischke
authors.forename: H-J
authors.initials: HJ
authors.lastname: Schuberth
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000097:blood* / Q000276:immunology*; D002460:Cell Line; D004283:Dog Diseases / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D004285:Dogs; D005434:Flow Cytometry / Q000379:methods / Q000662:veterinary*; D005455:Fluorescent Antibody Technique / Q000379:methods / Q000662:veterinary*; D006801:Humans; D009369:Neoplasms / Q000097:blood / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*
chemical_list: D001323:Autoantibodies