Fast multiplexed point-of-care diabetes autoantibody detection by homogeneous FRE

描述（由申请人提供）：本申请是一项床旁设备的可行性研究，用于快速多重分析最常见的自身抗体，这些抗体表明发生1型糖尿病（T1 D）的高风险，1型糖尿病是一种自身免疫性疾病，由产生胰岛素和调节身体能量代谢所需的细胞破坏引起。虽然与更常见的2型糖尿病分类，但出于实际目的，它们是基于病因学，诊断，代谢，流行病学和治疗的不同疾病。目前还没有已知的治愈方法，但人们对筛查和早期发现的兴趣正在增加，以便将来能够更好地控制和预防疾病。 在青年中，T1 D的新发病例率每年为每10，000人18至20例。在成人中，T1 D被认为占所有糖尿病诊断病例的5%，与2型糖尿病相比，1型糖尿病的成本高得不成比例。在怀孕前和怀孕的头三个月内控制不好的T1 D会导致5%到10%的孕妇出现严重的出生缺陷，15%到20%的孕妇会出现自然流产。 T1 D通常未被诊断，因为其症状难以确定，患者通常只在高血糖症发作后不久寻求急性症状的治疗。目前用于早期识别T1 D风险的最具特征的生物标志物是胰岛细胞自身抗体（伊卡），其不适当地靶向从垂死的胰腺细胞释放的细胞内蛋白。ICA不被认为是因果关系，而是对通过未检测到的T细胞杀死胰岛β细胞而释放到循环中的抗原的反应。换句话说，当因果疾病过程已经进行但无法检测时，ICA提供了一种循环生物标志物，以提醒临床医生T1 D风险升高。在 特别地，针对蛋白质胰岛素、IA-1、GAD 65和ZnT 8的自身抗体是高T1 D风险的标志，并且存在的ICA越多，风险越高。 第一阶段的可行性研究旨在开发一个快速的床旁诊断平台（EOSCAPE-T1 D），该平台可以同时测量医生办公室、实验室、诊所和医院中多种ICA的水平，只需最低限度的培训人员，几乎没有其他基础设施。目前的伊卡诊断是复杂的，需要长时间的等待，大量的实验室设备，受过专门训练的技术人员，往往放射性同位素。EOSCAPE-T1 D是一种封闭式检测盒，由申请组织的EOSCAPE-HIV平台改造而成，该平台开创了一套用于高性能核酸检测的新技术，每结果成本低。EOSCAPE-HIV于2009年9月被NIAID选择用于开发，用于在未来的HIV疫苗试验中区分真正的感染和疫苗诱导的血清阳性，目前正在进入开发的验证和确认阶段。EOSCAPE-T1 D将联合收割机专有的微流体致动器和检测盒结构与新型镧系元素时间分辨Forster共振能量转移检测相结合，用于T1 D自身抗体的均相多路检测。

DESCRIPTION (provided by applicant): This application is a feasibility study for a point of care device for rapid multiplexed analysis of the most common auto-antibodies that indicate high risk for developing type 1diabetes (T1D), an autoimmune disorder caused by destruction of the cells required to produce insulin and regulate the body's energy metabolism. While classified with the much more common type 2 diabetes, for practical purposes they are distinct diseases on the basis of etiology, diagnosis, metabolism, epidemiology, and treatment. There is no known cure, but interest is increasing in screening and early detection to enable better disease control and prevention in the future. Among youth the rate of new cases of T1D is between 18 and 20 per 10,000 each year. In adults, T1D is thought to account for as much as 5 percent of all diagnosed cases of diabetes, and the costs attributed to type 1diabetes compared to type 2 are disproportionately high. Poorly controlled T1D before conception and during the first trimester can cause major birth defects in 5 percent to 10 percent of pregnancies and spontaneous abortions in15 percent to 20 percent of pregnancies. T1D often goes undiagnosed because its symptoms are difficult to pinpoint, and patients generally only seek care for acute symptoms soon after the onset of hyperglycemia. The best characterized current biomarkers for early identification of T1D risk are islet cell auto-antibodies (ICA) which inappropriately target intracellular proteins released from dying pancreatic cells. ICAs are not thought to be causal but rather to be a response to antigens released into circulation by undetected T-cell killing of pancreatic islet beta cells. In other words, when the causal disease process is already underway but undetectable, ICAs provide a circulating biomarker to alert clinicians to elevated T1D risk. In particular, auto-antibodies against the proteins insulin, IA-1, GAD65, and ZnT8 are hallmarks of high T1D risk, and the more ICAs are present, the higher the risk. This Phase I feasibility study seeks to develop a fast point-of-care diagnostic platform (the EOSCAPE-T1D) which can simultaneously measure the levels of multiple ICAs in doctors¿ offices, labs, clinics, and hospitals with minimally trained staff and little to no other infrastructure. Current ICA diagnostis are complex, and require long wait times, extensive laboratory equipment, specially trained technicians, and often radioactive isotopes. The EOSCAPE-T1D is an enclosed cartridge point of care diagnostic adapted from the applicant organization's EOSCAPE-HIV platform, which pioneered a suite of new technologies for high-performance nucleic acid testing with a low cost-per-result. The EOSCAPE-HIV was chosen for development in September 2009 by NIAID for use in differentiating between true infection and vaccine-induced seropositivity in future HIV vaccine trials and is now entering the verification and validation stage of development. The EOSCAPE-T1D will combine proprietary micro-fluidic actuators and cartridge architecture with novel lanthanide time resolved Forster resonance energy transfer detection for a homogeneous multiplexed assay for T1D auto-antibodies.