3 Screen ELISA for High-Throughput Detection of Beta Cell Autoantibodies in Capillary Blood

Testing for beta cell autoantibodies is used for wide-scale identification of early stages of type 1 diabetes. This requires suitable screening assays. We aimed to establish screening that utilized a first step assay (3 Screen) able to detect autoantibodies to the target antigens glutamic acid decar...

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Published in:Diabetes technology & therapeutics Vol. 18; no. 11; p. 687
Main Authors: Ziegler, Anette-G, Haupt, Florian, Scholz, Marlon, Weininger, Katharina, Wittich, Susanne, Löbner, Stephanie, Matzke, Claudia, Gezginci, Cigdem, Riethausen, Stephanie, Beyerlein, Andreas, Zillmer, Stephanie, Amoroso, Marie, Coles, Rebecca, Powell, Michael, Furmaniak, Jadwiga, Smith, Bernard Rees, Winkler, Christiane, Bonifacio, Ezio, Achenbach, Peter
Format: Journal Article
Language:English
Published: United States 01.11.2016
Subjects:
Adolescent
Autoantibodies - blood
Capillaries
Cation Transport Proteins - immunology
Child
Diabetes Mellitus, Type 1 - immunology
Enzyme-Linked Immunosorbent Assay - methods
Female
Glutamate Decarboxylase - immunology
Humans
Insulin-Secreting Cells - immunology
Male
Receptor-Like Protein Tyrosine Phosphatases, Class 8 - immunology
Sensitivity and Specificity
Zinc Transporter 8
ISSN:1557-8593, 1557-8593
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Summary:Testing for beta cell autoantibodies is used for wide-scale identification of early stages of type 1 diabetes. This requires suitable screening assays. We aimed to establish screening that utilized a first step assay (3 Screen) able to detect autoantibodies to the target antigens glutamic acid decarboxylase-65 (GAD), insulinoma-associated antigen 2 (IA-2), and zinc transporter 8 (ZnT8) to identify children positive for multiple beta cell autoantibodies. An ELISA format was used where plates were coated with a mixture of recombinant GAD, IA-2, and ZnT8 W/R-dimer molecules. The performance was determined in venous blood from 686 first-degree relatives of patients with type 1 diabetes, and 200 patients at onset of type 1 diabetes, and applied as a screening assay in capillary blood from 33,639 general population children. The 3 Screen assay sensitivity for detecting autoantibody-positive patients at onset of type 1 diabetes was similar to that achieved by separate radiobinding assays (RBAs) for antibodies to GAD, IA-2, and ZnT8. Results in venous and capillary serum were correlated (R = 0.987). At a threshold corresponding to the 98th centile (29.1 U/mL) of all 33,639 capillary samples, the 3 Screen was positive in 123 samples with two or more RBA-positive antibodies to insulin, GAD, IA-2, or ZnT8, 146 with one antibody, and 479 that were RBA negative for beta cell autoantibodies. A 3 Screen ELISA was developed that was suitable for first step screening of multiple beta cell autoantibodies in capillary blood.
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ISSN:1557-8593
1557-8593
DOI:10.1089/dia.2016.0199