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

描述（由申请人提供）：该SBIR项目解决了用于检测与1型糖尿病（T1 D）相关的自身抗体的改进的基于新技术的测定的重要医学需求。T1 D影响了100万美国人，其发病率正以每年2%至5%的惊人速度增长。在T1 D中，针对胰岛细胞抗原的循环自身抗体反映了产生胰岛素的β细胞的自身免疫破坏过程。自身抗体在临床上是有用的，因为它们存在于免疫过程的早期，通常在患者发生高血糖症之前数年，并且可以用作标记物来识别T1 D风险个体并预测风险，特别是当存在针对多种抗原的自身抗体时。目前基于放射性免疫吸附（RIA）或酶联免疫吸附测定（ELISA）方法的测定法每次测定测试一种抗体，并且繁琐、昂贵，并且一些具有差的实验室间再现性。最近，一个单一的ELISA测定，用于同时检测两种抗体，但不能分配抗体的特异性变得可用。预防T1 D的新干预策略取决于T1 D前的早期检测，敏感和可靠的分析技术至关重要。基于我们的新型胶体技术的新型多重蛋白质微阵列检测的I期可行性研究已经非常成功，该检测可同时特异性检测三种自身抗体，即IAA、GAD-65和IA-2，这些抗体可为T1 D风险提供信息。在第二阶段，我们将开发用于临床诊断的“T1 D-自身免疫阵列测试系统”。完全集成的系统将包括一次性检测生物芯片，一个自动处理站，包括一个低成本的荧光阅读器，样本跟踪和分析软件。在第二阶段的成功结束时，我们将制定方案、程序和产品规范的基础，这些将成为“T1 D-自身免疫阵列检测系统”生产设计的基础，作为一种低成本、可靠、准确的检测产品，满足广泛的临床和研究环境的要求。公共卫生相关性：通过抗体检测预测T1 D对于正确识别风险受试者和风险量化至关重要。在T1 D中，尽管亲属中的风险较高，但90%的新病例是发生在没有既往家族史的家族中的散发病例。因此，在缺乏筛查人群的能力的情况下，大多数患有T1 D的儿童没有机会获得早期治疗，除非他们因为与患有这种疾病的人有关而接受筛查。显然，需要具有更好成本效益的新检测方法，以将自身抗体检测的益处扩展到大多数患有T1 D和其他自身免疫性疾病的个体。一种多重微阵列方法，将允许测试许多显着的T1 D自身抗体，也应该允许早期识别无症状的个人，并提供更好的机会，预防疾病。

DESCRIPTION (provided by applicant): This SBIR project addresses an important medical need for improved, new technology-based assays for detecting autoantibodies relevant to Type 1 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 autoantibodies 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 immune 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 autoantibodies against multiple antigens are present. Current assays, based on radio immunoadsorbant (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. Recently, a single ELISA assay that is used to detect two antibodies simultaneously but cannot assign antibody specificity became available. New intervention strategies for T1D prevention depend on early detection of pre-T1D and sensitive and reliable analytical techniques are crucial. The Phase I feasibility studies of a new multiplex protein microarray assay, based on our novel colloid technology to simultaneously and specifically detect three autoantibodies i.e. IAA, GAD-65 and IA-2 that are informative for T1D risk, have been very successful. In Phase II, we will develop the "T1D- Autoimmunity Array Testing System" for clinical diagnostic use. The fully integrated system will include disposable assay biochips, an automated processing station that includes a low cost fluorescence reader, sample tracking and analytical soft-ware. At the successful conclusion of Phase II, we will have developed protocols, procedures and the fundamentals of product specifications that will form the basis for production design of the "T1D- AutoimmunityArray Testing System" as a low cost, reliable, accurate testing product that meets the requirements of a broad range of clinical and research environments. PUBLIC HEALTH RELEVANCE: Prediction of T1D through antibody testing is critical for the proper identification of at-risk subjects and for risk quantification. In T1D, despite the higher risk in relatives, 90% of new cases are sporadic cases occurring in families with no prior family history. Thus, in the absence of the capability to screen populations at large, most children with T1D do not have the opportunity for early treatment unless they are screened because they are related to someone with the disease. Clearly, new assays with a better cost-benefit profile are needed to extend the benefit of autoantibody testing to the majority of individuals who develop T1D and other autoimmune diseases. A multiplex microarray methodology that would allow testing for numerous significant T1D autoantibodies should also allow for early identification of asymptomatic individuals and provide better chances of preventing disease.