Multiplex Microarray for the Detection of Pre-Type 1 Diabetes Autoantibodies

描述（由申请人提供）：这个SBIR项目解决了一个重要的医学需求，即改进的、基于新技术的检测与1型糖尿病（T1D）相关的自身抗体的方法。T1D影响着100万美国人，其发病率正以每年2%至5%的惊人速度增长。在T1D中，针对胰岛细胞抗原的循环自身抗体反映了自身免疫破坏产生胰岛素的β细胞的过程。自身抗体在临床上是有用的，因为它们在自身免疫过程的早期就存在，通常在患者出现高血糖前几年，并且可以用作识别T1D风险个体和预测风险的标记物，特别是当存在针对多种抗原的自身抗体时。目前基于放射免疫吸附剂（RIA）或酶联免疫吸附剂测定法（ELISA）的检测方法每次检测一种抗体，这些方法繁琐、昂贵，而且有些实验室间的可重复性很差。预防T1D的新干预策略依赖于早期发现T1D，而敏感可靠的分析技术至关重要。在第一阶段，我们将测试一种基于新型胶体技术的新型复合微阵列分析的可行性。将开发一种微阵列格式的免疫分析法，用于检测自身抗胰岛素（IAA）、抗胰岛素瘤相关抗原（IA-2）和抗谷氨酸脱羧酶（GAD65）这三种T1D自身抗体。单个微阵列检测将进行多路处理，并相对于当前的RIA和ELISA检测评估多路微阵列检测的性能。然后，我们将使用25个参考样本评估多重微阵列检测性能。第一阶段的结果将证明新型胶体复合微阵列技术作为支持检测T1D自身抗体的平台的可行性，并为第二阶段高效、准确、可靠的T1D临床诊断检测的全面开发和临床测试奠定基础。7. 通过自身抗体检测预测T1D对于正确识别有患此病风险的个体至关重要。在T1D中，尽管亲属风险较高，但90%的新病例是散发病例，发生在没有既往家族史的个体中。因此，由于缺乏对广大人群进行筛查的能力，大多数有T1D风险的个体没有被识别出来。显然，需要具有更好成本效益的新检测方法来扩大自身抗体检测的效益。所提出的多重微阵列方法是一种实用、高效的T1D自身抗体检测系统。这将有助于识别高危人群，从而为T1D预防研究提供宝贵的资源。

DESCRIPTION (provided by applicant): This SBIR project addresses an important medical need for improved, new technology-based assays for detecting auto antibodies relevant to Type I Diabetes (T1D). T1D affects a million Americans and its incidence is increasing at an alarming rate of 2 to 5% per year. In T1D, circulating auto antibodies against islet cell antigens reflect the process of autoimmune destruction of the insulin-producing beta-cells. Auto antibodies are clinically useful because they are present early in the autoimmune process, often years before patients develop hyperglycemia, and can be used as markers to identify individuals at risk for T1D and to predict risk, particularly when auto antibodies against multiple antigens, are present. Current assays, based on radio immuno adsorbant (RIA) or enzyme linked immunoadsorbant assays (ELISA) methods test for one antibody per assay and are cumbersome, expensive, and some have poor inter-laboratory reproducibility. New intervention strategies for T1D prevention depend on early detection of pre-T1D and sensitive and reliable analytical techniques are crucial. In Phase I, we will test the feasibility of a new multiplex microarray assay based on a novel colloid technology. A microarray format immunoassay will be developed for auto-anti-insulin (IAA), anti- insulinoma associated antigen ( IA-2) and anti-glutamic acid decarboxylase (GAD65)-three autoantibodies informative for T1D. The individual microarray assays will be multiplexed and the multiplex microarray assay performance evaluated relative to current RIA and ELISA assays. We will then evaluate the multiplex microarray assay performance using 25 reference samples. Results of Phase I will demonstrate the feasibility of the novel colloid multiplex micorarray technology as a platform for supporting assays to detect autoantibodies in T1D, and lay the groundwork for full scale development and clinical testing of an efficient, accurate, and reliable T1D clinical diagnostic assay in Phase II. 7. Project Narrative Prediction of T1D through autoantibody testing is critical for the proper identification of individuals at-risk for developing this disease. In T1D, despite the higher risk in relatives, 90% of new cases are sporadic cases occurring in individuals with no prior family history. Thus, in the absence of the capability to screen populations at large, most individuals at risk for T1D are not identified. Clearly, new assays with a better cost-benefit profile are needed to expand the benefit of autoantibody testing. The proposed multiplex microarray methodology is a practical, efficient system to test for T1D autoantibodies. This should allow for the identification of at risk individuals and thus provide a valuable resource for T1D prevention studies.