Digital Microfluidics - Minimizing blood volume for pediatric coagulation screening

描述(申请人提供)：据报道，在接受心脏手术的新生儿中，高达40%遭受重大血栓形成事件。因先天或后天高凝状态而增加血栓形成风险的儿科患者需要个体化的抗凝治疗。亲血栓症的存在会影响抗凝时间和持续时间的临床决策，特别是在静脉血栓形成的儿童。治疗的持续时间取决于危险因素的数量和血栓形成情况。高凝小组筛查可为治疗管理提供风险分层。高凝状态筛查适用于有血栓家族史的儿科患者，或遭受重大血栓形成或血栓栓塞症事件的儿童患者。然而，大量患者在没有达到目前的筛查标准的情况下发生血栓形成。目前可用的筛查测试昂贵、耗时，而且需要单独订购，需要大量血液样本。这些问题使得常规的全面高凝筛查在临床上是不切实际的，特别是在儿科患者中。因此，迫切需要一种廉价的高凝实验室面板，这种面板需要非常少量的血液，具有快速的周转时间，并且可以应用于更多的潜在风险患者。这样的诊断面板可以通过采用数字微流控系统来实现。对数字微流控芯片进行的高凝测试将实现便携式自动化、快速周转时间和多种分析，包括不同类型的分析(免疫分析和功能分析)，每个测试使用大约100个NL样本。一种使用数字微流控技术进行高凝状态面板测试的一次性盒 将开发一系列液滴。最终目标是开发一种单一平台，可以使用手指刺伤全血样本进行多重免疫分析或酶联免疫吸附分析(ELISA)和功能分析，以确定高凝状态的风险，最初的重点是接受此类测试的儿科患者。在第一阶段，我们建议将多重免疫分析(抗凝血酶III、蛋白C和纤溶酶原激活物抑制物-1)与功能分析(抗凝血酶III、蛋白C、纤溶酶原、FVIII和凝血酶生成)整合在一个一次性试剂盒上，从而实现完整的血栓亲和性面板。在第二阶段，将开展试剂盒上试剂储存、全面质量控制以及产品商业化的其他必要活动。

DESCRIPTION (provided by applicant): It is reported that up to 40% of neonates undergoing cardiac surgery suffer a major thrombosis event. Pediatric patients who are at increased risk for thrombosis due to either congenital or acquired hypercoagulability require individualized anticoagulation management. Presence of a thrombophilia influences clinical decision making regarding timing and duration of anticoagulation, particularly in children with venous thrombosis. The duration of therapy is guided by the number of risk factors and thrombophilic conditions. Hypercoagulability panel screening allows risk stratification for therapeutic management. Screening for hypercoagulability is indicated in pediatric patients with a family history of thrombosis, or in those who have suffered a major thrombotic or thromboembolic event. However, a large number of patients develop thrombosis without meeting current criteria for screening. Currently available screening tests are expensive, time-consuming and ordered individually, requiring large volumes of blood sample. These issues make routine screening for a full hypercoagulability panel clinically impractical, especially in pediatric patients. Hence, thre is a compelling need for an inexpensive hypercoagulability laboratory panel that requires very small volumes of blood, has a rapid turn-around time and can be applied to a larger number of potentially at-risk patients. Such a diagnostic panel is possible by employing a digital microfluidic system. Hypercoagulability tests on a digital microfluidic cartridge will enable portable automation, rapid turn-around time, and multiplexing several assays, including different types of assays (immunoassays and functional) utilizing about 100 nL sample per test. A disposable cartridge for hypercoagulability panel testing using digital microfluidic manipulation of droplets will be developed. The ultimate goal is the development of a single platform that can perform multiplexed immunoassays or enzyme-linked immunosorbent assay (ELISA) and functional assays using a finger prick whole blood sample to determine hypercoagulability risk, with initial emphasis on pediatric patients undergoing such testing. In Phase I, we propose to integrate the multiplexed immunoassays (Anti-thrombin III, Protein C and Plasminogen Activator Inhibitor-1) with functional assays (Anti-thrombin III, Protein C, Plasminogen, FVIII and Thrombin generation) allowing for a complete thrombophilia panel on a single disposable cartridge. In Phase II, on-cartridge reagent storage, comprehensive quality control, and other necessary activities for commercialization of a product will be developed.