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

Diabetes Technol Ther. 2016 Nov;18(11):687-693. doi: 10.1089/dia.2016.0199. Epub 2016 Aug 23.


Background: 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.

Methods: 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.

Results: 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.

Conclusion: A 3 Screen ELISA was developed that was suitable for first step screening of multiple beta cell autoantibodies in capillary blood.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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


  • Autoantibodies
  • Cation Transport Proteins
  • SLC30A8 protein, human
  • Zinc Transporter 8
  • PTPRN protein, human
  • Receptor-Like Protein Tyrosine Phosphatases, Class 8
  • Glutamate Decarboxylase