Commisariat à l'Energie Atomique, iBiTec-S, Service de Pharmacologie et d'Immunoanalyse, Commisariat à l'Energie Atomique Saclay, Gif sur Yvette, France.
PLoS One. 2013 Jul 29;8(7):e69021. doi: 10.1371/journal.pone.0069021. Print 2013.
Type 1 diabetes (T1D) is an autoimmune disease which results from the destruction of pancreatic beta cells. Autoantibodies directed against islet antigens are valuable diagnostic tools. Insulin autoantibodies (IAAs) are usually the first to appear and also the most difficult to detect amongst the four major islet autoantibodies. A non-radioactive IAA bridging ELISA was developed to this end. In this assay, one site of the IAAs from serum samples is bound to a hapten-labeled insulin (GC300-insulin), which is subsequently captured on anti-GC300 antibody-coated 96-well plates. The other site of the IAAs is bound to biotinylated insulin, allowing the complex to be detected by an enzyme-streptavidin conjugate. In the present study, 50 serum samples from patients with newly diagnosed T1D and 100 control sera from non-diabetic individuals were analyzed with our new assay and the results were correlated with an IAA radioimmunoassay (RIA). Using IAA bridging ELISA, IAAs were detected in 32 out of 50 T1D children, whereas with IAA RIA, 41 out of 50 children with newly diagnosed T1D were scored as positive. In conclusion, the IAA bridging ELISA could serve as an attractive approach for rapid and automated detection of IAAs in T1D patients for diagnostic purposes.
1 型糖尿病（T1D）是一种自身免疫性疾病，由胰腺β细胞破坏引起。针对胰岛抗原的自身抗体是有价值的诊断工具。胰岛素自身抗体（IAAs）通常是最早出现的，也是四种主要胰岛自身抗体中最难检测的。为此，开发了一种非放射性 IAA 桥接 ELISA。在该测定法中，血清样本中的 IAAs 的一个位点与半抗原标记的胰岛素（GC300-胰岛素）结合，随后将其捕获在抗 GC300 抗体包被的 96 孔板上。IAAs 的另一个位点与生物素化胰岛素结合，允许通过酶-链霉亲和素缀合物检测复合物。在本研究中，用我们的新测定法分析了 50 例新诊断的 T1D 患者和 100 例非糖尿病个体的血清样本，并将结果与 IAA 放射免疫测定（RIA）相关联。使用 IAA 桥接 ELISA，在 50 例 T1D 儿童中有 32 例检测到 IAAs，而使用 IAA RIA，在 50 例新诊断的 T1D 儿童中有 41 例被评为阳性。总之，IAA 桥接 ELISA 可作为一种有吸引力的方法，用于快速和自动检测 T1D 患者的 IAAs，用于诊断目的。
