title: Next-Generation Autoantibody Testing by Combination of Screening and Confirmation-the CytoBead® Technology.
abstract: Occurrence of autoantibodies (autoAbs) is a hallmark of autoimmune diseases, and the analysis thereof is an essential part in the diagnosis of organ-specific autoimmune and systemic autoimmune rheumatic diseases (SARD), especially connective tissue diseases (CTDs). Due to the appearance of autoAb profiles in SARD patients and the complexity of the corresponding serological diagnosis, different diagnostic strategies have been suggested for appropriate autoAb testing. Thus, evolving assay techniques and the continuous discovery of novel autoantigens have greatly influenced the development of these strategies. Antinuclear antibody (ANA) analysis by indirect immunofluorescence (IIF) on tissue and later cellular substrates was one of the first tests introduced into clinical routine and is still an indispensable tool for CTD serology. Thus, screening for ANA by IIF is recommended to be followed by confirmatory testing of positive findings employing different assay techniques. Given the continuous growth in the demand for autoAb testing, IIF has been challenged as the standard method for ANA and other autoAb analyses due to lacking automation, standardization, modern data management, and human bias in IIF pattern interpretation. To address these limitations of autoAb testing, the CytoBead® technique has been introduced recently which enables automated interpretation of cell-based IIF and quantitative autoAb multiplexing by addressable microbead immunoassays in one reaction environment. Thus, autoAb screening and confirmatory testing can be combined for the first time. The present review discusses the history of autoAb assay techniques in this context and gives an overview and outlook of the recent progress in emerging technologies.
authors.affiliation: GA Generic Assays GmbH, Dahlewitz, Berlin, Germany.
authors.forename: Mandy
authors.identifier: http://orcid.org/0000-0002-8660-4109
authors.initials: M
authors.lastname: Sowa
authors.affiliation: Institute of Biotechnology, Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Senftenberg, Germany.
authors.forename: Rico
authors.initials: R
authors.lastname: Hiemann
authors.affiliation: Institute of Biotechnology, Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Senftenberg, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Schierack
authors.affiliation: Institute of Molecular and Clinical Immunology, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Reinhold
authors.affiliation: Institute of Immunology, Medical Faculty, Technical University Dresden, Dresden, Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: Conrad
authors.affiliation: GA Generic Assays GmbH, Dahlewitz, Berlin, Germany. dirk.roggenbuck@b-tu.de.|Institute of Biotechnology, Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Senftenberg, Germany. dirk.roggenbuck@b-tu.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Roggenbuck
mesh_terms: D000974:Antibodies, Antinuclear / Q000097:blood / Q000276:immunology; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D001327:Autoimmune Diseases / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D015551:Autoimmunity; D015415:Biomarkers; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D006801:Humans
chemical_list: D000974:Antibodies, Antinuclear; D001323:Autoantibodies; D015415:Biomarkers
keywords: Autoimmune disease; Digital fluorescence; Indirect immunofluorescence; Multiplex diagnostics; Second-generation autoantibody testing