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

描述(申请人提供)：本申请是一种护理设备的可行性研究，用于对最常见的自身抗体进行快速多路分析，这些抗体表明患1型糖尿病(T1D)的风险很高，T1D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被破坏而引起。虽然被归类为更常见的2型糖尿病，但实际上它们是根据病因、诊断、新陈代谢、流行病学和治疗而不同的疾病。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越来越大，以便在未来更好地控制和预防疾病。在年轻人中，每年T1D新发病例的比例在每万人中有18到20人。在成年人中，T1D被认为占所有确诊糖尿病病例的5%，与2型糖尿病相比，1型糖尿病的成本高得不成比例。在怀孕前和怀孕前三个月控制不好的T1D会导致5%到10%的妊娠出现严重的出生缺陷，15%到20%的怀孕会导致自然流产。T1D经常被漏诊，因为它的症状很难确定，而且患者通常只在高血糖发作后不久就寻求急性症状的治疗。目前用于早期识别T1D风险的最具特征性的生物标记物是胰岛细胞自身抗体(ICA)，它不适当地靶向从死亡的胰腺细胞释放的细胞内蛋白。ICAS被认为不是因果的，而是对未被检测到的T细胞对胰岛β细胞的杀伤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开始但无法检测时，ICAS提供了一个循环生物标志物，以提醒临床医生T1D风险增加。在……里面 特别是，针对胰岛素、IA-1、GAD65和ZnT8蛋白的自身抗体是T1D风险高的标志，ICA越多，风险越高。这项第一阶段的可行性研究旨在开发一个快速的医疗保健点诊断平台(EOSCAPE-T1D)，该平台可以同时测量医生办公室、实验室、诊所和医院中多个ICA的水平，这些医院的工作人员只受过最低程度的培训，几乎没有其他基础设施。目前的ICA诊断很复杂，需要很长的等待时间，需要大量的实验室设备，需要经过专门培训的技术人员，而且通常还需要放射性同位素。EOSCAPE-T1D是一种封闭式试剂盒，改编自申请组织的EOSCAPE-HIV平台，该平台开创了一套高性能核酸检测的新技术，每结果成本较低。NIAID于2009年9月选择EOSCAPE-HIV进行开发，用于在未来的艾滋病毒疫苗试验中区分真实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-T1D将专有的微流体致动器和盒结构与新颖的稀土元素时间分辨Forster共振能量转移检测相结合，用于T1D自身抗体的均质多路复用检测。

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.