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 .
Diabetes Technol Ther. 2016 Nov;18(11):687-693. doi: 10.1089/dia.2016.0199. Epub 2016 Aug 23.
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 ZnT8W/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.
检测β细胞自身抗体用于广泛识别 1 型糖尿病的早期阶段。这需要合适的筛查检测。我们旨在建立一种筛选方法，该方法使用第一步检测（3 筛）能够检测谷氨酸脱羧酶-65（GAD）、胰岛相关抗原 2（IA-2）和锌转运蛋白 8（ZnT8）的自身抗体，以识别多个β细胞自身抗体阳性的儿童。
使用 ELISA 格式，其中板用重组 GAD、IA-2 和 ZnT8W/R-二聚体分子的混合物包被。在 686 名 1 型糖尿病患者的一级亲属和 200 名 1 型糖尿病发病患者的静脉血中测定性能，并在 33639 名普通人群儿童的毛细血管血中作为筛查检测进行应用。
3 筛检测在检测 1 型糖尿病发病患者自身抗体阳性患者的敏感性与单独用于检测 GAD、IA-2 和 ZnT8 抗体的放射结合测定（RBA）相似。静脉和毛细血管血清的结果相关（R=0.987）。在对应于所有 33639 个毛细血管样本第 98 百分位数（29.1 U/mL）的阈值下，3 筛在 123 个样本中呈阳性，这些样本具有两个或更多胰岛素、GAD、IA-2 或 ZnT8 的 RBA 阳性抗体，146 个样本具有一个抗体，479 个样本 RBA 阴性β细胞自身抗体。
开发了一种 3 筛 ELISA，适用于毛细血管血中多种β细胞自身抗体的第一步筛选。
