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

描述(由申请人提供)：该SBIR项目满足了对检测与1型糖尿病(T1D)相关的自身抗体的改进的、基于新技术的分析的重要医学需求。T1D影响着一百万美国人，其发病率正以每年2%至5%的惊人速度增长。在T1D中，循环中的针对胰岛细胞抗原的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。自身抗体在临床上很有用，因为它们在免疫过程的早期就出现了，通常是在患者出现高血糖的前几年，可以作为标志物来识别T1D的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。目前基于放射免疫吸附(RIA)或酶联免疫吸附分析(EL ISA)方法的检测方法每次检测一种抗体，且繁琐、昂贵，而且一些实验室间重复性较差。最近，一种同时检测两种抗体但不能指定抗体特异性的单一ELISA法成为可能。预防T1D的新干预策略依赖于T1D前的早期检测，敏感和可靠的分析技术至关重要。基于我们的新胶体技术，同时特异性地检测三种与T1D风险有关的自身抗体，即IAA，GAD-65和IA-2，一种新的多重蛋白质微阵列分析的第一阶段可行性研究非常成功。在第二阶段，我们将开发用于临床诊断的T1D-自身免疫阵列检测系统。完全集成的系统将包括一次性检测生物芯片、一个自动化处理站，其中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，我们将制定协议、程序和产品规格的基本原则，为“T1D-自动免疫阵列测试系统”的生产设计奠定基础，作为一种低成本、可靠、准确的测试产品，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T1D对于正确识别高危受试者和风险量化至关重要。在T1D中，尽管亲属的风险较高，但90%的新病例是发生在没有家族史的家庭中的零星病例。因此，在缺乏对广大人群进行筛查的能力的情况下，大多数患有T1D的儿童没有机会得到早期治疗，除非他们接受筛查，因为他们与该疾病的某个人有亲属关系。显然，为了将自身抗体检测的好处扩大到大多数患有T1D和其他自身免疫性疾病的人，需要具有更好成本效益的新检测方法。可以检测大量重要的T1D自身抗体的多重微阵列方法也应该能够及早识别无症状的个体，并提供更好的预防疾病的机会。

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.