A rapid microfluidic diagnostic assay for the measurement of Direct Oral Anticoagulants from patient whole blood

摘要：直接口服抗凝剂(DOAC)正成为首选的治疗方法 抗凝，尤其是在溶栓性疾病中。预计下一年全球DOAC使用量将达到300亿美元 五年，超过华法林等其他抗凝血剂。虽然DOAC非常有效，但它可以增加出血 风险，并使紧急护理复杂化，其中可能包括使用昂贵的DOAC逆转剂。当前状态 诊断无法充分识别DOAC引起的问题，因此存在越来越多的安全问题 联合委员会呼吁进行实验室测试监测的药物类别，以及证据- 基于安全DOAC和反转剂使用的方法。无论是治疗中风/VTE/PE患者，还是心脏 心律失常患者，或创伤/紧急手术患者，实时确定患者的状态 抗凝，包括DOAC型(凝血因子Xa抑制物与直接凝血酶抑制物)对快速护理至关重要 避免不良出血的决定。核心实验室检测DOAC的一个重要限制是长度 到结果的时间，这可能会超过典型的20分钟决策窗口。FloBio是 开发首个DOAC体外诊断产品，为医生提供快速诊断知识 急救环境中的抗凝状态。使用我们的分析仪/成像站和一次性DOAC检测 盒/微流控装置，我们的新方法结合了血流动力学和离散的凝块激活，以 模拟生理性的血液凝结。在不到10分钟的时间里，医疗保健提供者将知道患者的相对DOAC 专注度和DOAC分类。 FloBio的第一阶段工作已经展示了一种可扩展、多路、一次性DOAC试剂盒/微流控 促进体外自然凝血级联的装置，用于测量分析信号的仪器， 分析软件，为最终用户提供简单的结果评估和验证的临床数据 该方法和装置用于检测人体血液中DOAC的种类和水平的可行性。FloBio的创新是一种 这将是市场上第一个提供纤维蛋白和血小板聚集的实时数据的诊断分析 在基础血管和凝血蛋白上，在流动下，重现体内的凝血过程。 在第二阶段，FloBio将专注于在加州大学进行的100名受试者的临床研究中验证DOAC分析。 宾夕法尼亚州。这项研究将验证该方法检测类别和DOAC水平的能力，在 临床患者群体。使用DOAC添加健康成人血液的第二项临床研究将确定 仪器、化验和系统范围、灵敏度、特异度、方差和一致性。临床化验数据将 对照检测的黄金标准--LCMS进行了验证。关键化验功能的自动化也将是 评估以减少分析中的差异，以及完全集成、自动化、一次性DOAC的可行性 试剂盒将在健康的人体血液样本中进行验证。这项工作的结果将是一个 经过审查的DOAC检测系统，已准备好在IIb阶段全面开发。

Abstract: Direct oral anticoagulants (DOACs) are becoming the preferred approach to managing anticoagulation, particularly in thrombolytic disorders. DOAC use is anticipated to reach $30B globally in the next five years, overtaking other anticoagulants such as warfarin. While highly effective, DOACs can elevate bleeding risks, and complicate emergency care, which may include the use of expensive DOAC reversal agents. As current diagnostics fail to adequately identify problems caused by DOACs, there exists growing safety concerns for this class of drugs that are reinforced by the Joint Commission’s call for laboratory test monitoring, and evidence- based approaches for safe DOAC, and reversal agent use. Whether treating a stroke/VTE/PE patient, heart arrhythmia patient, or trauma/urgent surgery patient, the real-time determination of a patient’s state of anticoagulation, including DOAC type (factor Xa inhibitor vs. direct thrombin inhibitor) is critical for rapid care decisions to avoid adverse bleeding. A significant limitation of core laboratory detection of DOACs is the length of time to results, which can exceed more than the typical 20-minute decision making window. FloBio is developing the first DOAC in vitro diagnostic product to provide physicians with rapid knowledge of anticoagulation state in emergency care settings. With our analyzer/imaging station, and disposable DOAC assay cartridge/microfluidic device, our novel approach combines hemodynamic flow, and discrete clot activation, to mimic physiological blood clotting. In less than 10 minutes, a healthcare provider will know a patient’s relative DOAC concentration, and the DOAC classification. FloBio’s Phase I work has demonstrated a scalable, multiplex, disposable DOAC assay cartridge/microfluidic device that facilitates the natural blood clotting cascade in vitro, an instrument for measuring assay signals, analytic software to provide the end user with a simple outcomes assessment, and clinical data that validates the feasibility of the assay and device to detect DOAC class and level in human blood. FloBio’s innovation is a diagnostic assay that will be the first in the market to provide real-time data on fibrin and platelet accumulation on basal vascular and coagulation proteins, under flow, that recapitulates in vivo coagulation processes. In Phase II, FloBio will focus on validating the DOAC assay in a 100 subject clinical study at the University of Pennsylvania. This study will validate the capability of the assay to detect both class and DOAC level, within a clinical patient population. A second clinical study with DOAC spiked healthy adult blood will determine the device, assay and system range, sensitivity, specificity, variance, and consistency. Clinical assay data will be validated against the gold standard of detection, LCMS. Automation of critical assay functions will also be evaluated to reduce variation in the assay, and the feasibility of a fully integrated, automatable, disposable DOAC assay cartridge will be demonstrated in healthy human blood samples. The outcome of the work will be a fully vetted DOAC assay system that is ready for full development in Phase IIb.