title: 3 Screen ELISA for High-Throughput Detection of Beta Cell Autoantibodies in Capillary Blood.
abstract: 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 ZnT8325W/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.
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .|2 Forschergruppe Diabetes e.V. , Neuherberg, Germany .
authors.forename: Anette-G
authors.initials: AG
authors.lastname: Ziegler
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Florian
authors.initials: F
authors.lastname: Haupt
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Marlon
authors.initials: M
authors.lastname: Scholz
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Katharina
authors.initials: K
authors.lastname: Weininger
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Susanne
authors.initials: S
authors.lastname: Wittich
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Stephanie
authors.initials: S
authors.lastname: Löbner
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Claudia
authors.initials: C
authors.lastname: Matzke
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Cigdem
authors.initials: C
authors.lastname: Gezginci
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Stephanie
authors.initials: S
authors.lastname: Riethausen
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Andreas
authors.initials: A
authors.lastname: Beyerlein
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .
authors.forename: Stephanie
authors.initials: S
authors.lastname: Zillmer
authors.affiliation: 3 FIRS Laboratories , RSR Ltd., Parc Ty Glas, Llanishen, Cardiff, United Kingdom .
authors.forename: Marie
authors.initials: M
authors.lastname: Amoroso
authors.affiliation: 3 FIRS Laboratories , RSR Ltd., Parc Ty Glas, Llanishen, Cardiff, United Kingdom .
authors.forename: Rebecca
authors.initials: R
authors.lastname: Coles
authors.affiliation: 3 FIRS Laboratories , RSR Ltd., Parc Ty Glas, Llanishen, Cardiff, United Kingdom .
authors.forename: Michael
authors.initials: M
authors.lastname: Powell
authors.affiliation: 3 FIRS Laboratories , RSR Ltd., Parc Ty Glas, Llanishen, Cardiff, United Kingdom .
authors.forename: Jadwiga
authors.initials: J
authors.lastname: Furmaniak
authors.affiliation: 3 FIRS Laboratories , RSR Ltd., Parc Ty Glas, Llanishen, Cardiff, United Kingdom .
authors.forename: Bernard Rees
authors.initials: BR
authors.lastname: Smith
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .|2 Forschergruppe Diabetes e.V. , Neuherberg, Germany .
authors.forename: Christiane
authors.initials: C
authors.lastname: Winkler
authors.affiliation: 4 Centre for Regenerative Therapies Dresden, Faculty of Medicine, Technische Universität , Dresden, Germany .|5 Paul Langerhans Institute Dresden of Helmholtz Centre Munich at University Clinic Carl Gustav Carus of Technische Universität , Dresden, Germany .
authors.forename: Ezio
authors.initials: E
authors.lastname: Bonifacio
authors.affiliation: 1 Institute of Diabetes Research, Helmholtz Zentrum München, and Forschergruppe Diabetes, Klinikum rechts der Isar, Technische Universität München , Neuherberg, Germany .|2 Forschergruppe Diabetes e.V. , Neuherberg, Germany .
authors.forename: Peter
authors.initials: P
authors.lastname: Achenbach
mesh_terms: D000293:Adolescent; D001323:Autoantibodies / Q000097:blood*; D002196:Capillaries; D027682:Cation Transport Proteins / Q000276:immunology; D002648:Child; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005968:Glutamate Decarboxylase / Q000276:immunology; D006801:Humans; D050417:Insulin-Secreting Cells / Q000276:immunology*; D008297:Male; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8 / Q000276:immunology; D012680:Sensitivity and Specificity; D000075703:Zinc Transporter 8
chemical_list: D001323:Autoantibodies; D027682:Cation Transport Proteins; C492522:SLC30A8 protein, human; D000075703:Zinc Transporter 8; C516862:PTPRN protein, human; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D005968:Glutamate Decarboxylase