Development of An Automated High-Throughput Dried Blood Spot Assay to Facilitate Large Scale Screening for Type 1 Diabetes Risk

开发自动化高通量干血斑测定法以促进大规模筛查 1 型糖尿病风险

• 批准号: 10020787
• 负责人: David Seftel
• 金额: $ 74.12万
• 依托单位: ENABLE BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-19 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10020787
• 关键词: Accident and Emergency department; Achievement; Address; Adult; Affect; Agglutination; Agreement; American; Antibodies; Autoantibodies; Autoimmune Diseases; Automation; Biological Assay; Biological Sciences; Blinded; Blood; Blood Screening; Child; Classification; Clinical; Clinical Laboratory Information Systems; Cold Chains; Collaborations; Collection; Communities; Custom; Data; Decentralization; Detection; Development; Devices; Diabetes autoantibodies; Diabetic Ketoacidosis; Diagnosis; Disease; Economic Burden; Effectiveness; Fingers; General Population; Gold; Immunoassay; Incidence; Individual; Information Systems; Insulin Antibodies; Insulin-Dependent Diabetes Mellitus; Islet Cell; Laboratories; Life; Liquid substance; Manuals; Measures; Methods; Modification; Paper; Patients; Performance; Phase; Preparation; Procedures; Process; Protocols documentation; Quality of life; Radioimmunoassay; Readiness; Reagent; Research Personnel; Resolution; Risk; Robotics; Sampling; Serum; Shipping; Signal Transduction; Solid; Spottings; Standardization; Sudden Death; System; Testing; Universities; Validation; Venous blood sampling; base; chronic autoimmune disease; cloud based; cost; cost effective; design; diabetes risk; improved; islet; medical schools; population based; prevent; program costs; programs; quality assurance; robotic system; screening; screening program; young adult

Type 1 diabetes (T1D) is a chronic autoimmune disease that affects millions of children and adults in the US. Up to 40% of T1D patients are diagnosed in the emergency room with life-threatening diabetic ketoacidosis (DKA), resulting in severe clinical complications and a substantial economic burden. Programs that screen for blood-borne markers of early-stage T1D (anti-islet cell autoantibodies) can significantly diminish the incidence of DKA and improve patient quality of life. However, large-scale implementation of such screening campaigns is hindered by the high cost and low throughput of radioimmunoassay (RIA), the gold-standard method for measuring anti-islet autoantibodies. Moreover, a significant portion of screening program costs are associated with phlebotomy, cold-chain storage and shipping. To address this, Enable Biosciences aims to develop a cost-effective, multiplex, high- throughput and non-radioactive immunoassay to diagnose T1D and screen for individuals at risk for T1D using low-cost easy-to-collect dried blood spots (DBS). The successful development of this high- throughput dried blood spot T1D assay can substantially increase the effectiveness of general population- based T1D screening programs. In a JDRF- and Stanford-supported initiative, we were gratified that a serum-based Enable assay achieved the highest performance for anti-GAD, anti-IA2 and a close second-to-highest performance for anti-insulin antibodies in the recent blinded 2018 Islet-cell Autoantibody Standardization Program (IASP) competition out of 50 participating laboratories. The IASP data were generated with an automated Enable serum assay using a custom-made Enable analyzer created in collaboration with Hamilton Robotics. Furthermore, we showed that our manual DBS- based Enable assay correlated strongly with a serum-based assay (R=0.90-0.96). The concordance as evaluated by Cohen’s test showed kappa between 0.85-0.89. The positive and negative agreements for all three aforementioned autoantibodies range from 85%-99%. Notably, we also demonstrated less than 10% degradation of assay signals when DBS cards were stored at 37°C for 4 weeks. Building on these solid technical achievements, which we consider as our “Phase I-like Results” described in detail in this proposal, we aim to further automate our DBS assay procedures for high-throughput functionality including DBS elution and DBS eluent testing using diverse samples, such as those from Stanford University School of Medicine. We expect to deliver an integrated DBS T1D autoantibody assay (elution/testing automation device and reagent kits) to serve large screening centers and CLIA- laboratories. We further plan to obtain CE Mark classification and FDA approval to further decentralize DBS T1D autoantibody testing to be able to address the important needs of communities, clinicians and researchers worldwide.

1型糖尿病（T1D）是一种慢性自身免疫性疾病，影响数百万儿童和成人 我们。在急诊室诊断出多达40％的T1D患者患有威胁生命的糖尿病 酮症酸中毒（DKA），导致严重的临床并发症和实质性的经济负担。 筛选早期T1D的血传播标记（抗ISLET细胞自动抗体）的程序可以 大大降低了DKA的事件并提高患者的生活质量。但是，大规模 高成本和低吞吐量阻碍了此类筛选活动的实施 放射免疫测定（RIA），用于测量抗ISLET自身抗体的金标准方法。而且， 筛选计划的大部分成本与静脉切开术，冷链存储和 船运。为了解决这个问题，使生物科学旨在开发一种成本效益，多重，高级 吞吐量和非放射性免疫测定法，以诊断有T1D风险的个体T1D和屏幕 使用低成本易于收集的干血点（DBS）。这个高级的成功发展 吞吐量干血点T1D分析可以大大提高一般人群的有效性 - 基于T1D筛选程序。在JDRF和斯坦福支持的倡议中，我们被评为 基于血清的启用测定实现了抗GAD，抗IA2和关闭的最高性能 最近盲人2018 Islet细胞中抗胰岛素抗体的二到最高性能 在50个参与实验室中，自动抗体标准化计划（IASP）竞争。 使用定制启用的自动化启用血清分析生成IASP数据 分析仪与汉密尔顿机器人技术合作创建。此外，我们证明了我们的手册DBS- 基于启用测定与基于血清的测定密切相关（r = 0.90-0.96）。一致性 通过Cohen的测试评估，kappa在0.85-0.89之间。正面和负面协议 所有关于自身抗体的三个三者范围为85％-99％。值得注意的是，我们还表现得比 当将DBS卡在37°C下储存4周时，测定信号的降解为10％。建立在这些基础上 扎实的技术成就，我们认为这是我们的“类似I期结果”的详细描述 提案，我们旨在进一步自动化DBS分析程序的高通量功能 包括使用潜水员样品（例如斯坦福大学的DBS洗脱和DBS洗脱液测试） 大学医学院。我们希望提供集成的DBS T1D自动抗体测定法 （洗脱/测试自动化设备和试剂套件）用于大型筛选中心和Clia- 实验室。我们进一步计划获得CE Mark分类和FDA批准，以进一步分散 DBS T1D自动抗体测试能够满足社区，临床医生和 全球研究人员。