Digital Microfluidics - Minimizing Blood Volume for Pediatric Coagulation Screening

摘要 数字微流控技术-最大限度地减少儿科凝血筛查的血容量 (SBIR IIB期） 接受心脏手术的新生儿和幼儿发生重大心脏病的风险显著增加。 血栓形成事件和全面的高凝状态筛查适用于患有 严重血栓形成或血栓栓塞事件，或有血栓形成家族史的患者。当前可用 凝血筛查测试是昂贵的，耗时的，必须单独订购，这增加了 所需血液样本的累积体积。这些问题使得筛选和监测成为一个全面的问题。 高凝状态面板临床上不切实际，特别是在新生儿和高危儿科患者， 心脏手术因此，迫切需要一种廉价的高凝性测试板，其需要 非常小的血液体积，具有快速的周转时间，并且具有应用于更大数量的潜力 在手术管理过程中的多个时间点， 在该SBIR项目（HL 125484）的第I和第II阶段，我们开发了一组五个关键凝血 在我们的近患者数字微流体仪器上进行分析。该测定包括针对血管性血友病的抗原测定 因子（VWF）和蛋白S，加上抗凝血酶III（ATIII）、蛋白C和因子VIII的活性测定。所有5 从一滴全血样品（< 50 μ L）共同进行测定。IIB阶段的额外资金将 使我们能够开发用于凝血因子V Leiden和凝血酶原（凝血因子II）遗传学的互补分子测定法， 突变，这将在相同的测定平台上自动化。这些分子检测的增加将 提供更全面的凝血风险评估，并将提高我们的商业潜力， 最终产品。我们还将进行CLSI指导的完整的分析和临床验证， 凝血试验和准备510（k）FDA批准的监管申请。目前没有一个单一的， 市场上的小容量检测平台，可支持抗原、活性和分子的快速检测 分析物，以提供高凝风险的全面评估。我们的测试平台和分析 经过专门设计，可满足快速、低复杂性测试的技术要求， 样本量和解决儿科凝血管理中关键的未满足需求。 我们的方法通过与我们在波士顿儿童医院的临床合作伙伴的合作得到了加强， 测试的潜在临床影响，数字微流体技术在全血测试中的新应用 在患者附近，并通过彻底的市场分析和其他战略营销，已启动的 产品通过其他资金来源。数字微流控分析将适合在患者附近使用 最低的培训要求。我们的测试的主要预期用户将是护士或实验室技术人员， 儿科护理环境，但我们预计该系统可以很容易地扩展到其他市场，如 成人心脏病患者和农村或资源有限的医院环境，目前无法执行 完整的凝血试验面板。

ABSTRACT Digital Microfluidics – Minimizing Blood Volume for Pediatric Coagulation Screening (SBIR Phase IIB) Newborns and young children undergoing cardiac surgery are at significantly increased risk for a major thrombosis event, and comprehensive hypercoagulability screening is indicated in children who have suffered a major thrombotic or thromboembolic event, or who have a family history of thrombosis. Currently available coagulation screening tests are expensive, time-consuming, and must be ordered individually, which increases the cumulative volume of blood sample required. These issues make screening and monitoring for a full hypercoagulability panel clinically impractical, especially in newborns and high-risk pediatric patients undergoing cardiac surgery. Hence, there is a compelling need for an inexpensive hypercoagulability test panel that requires very small volumes of blood, has a rapid turn-around time, and has the potential to be applied to a larger number of at-risk patients at multiple time points during their surgical management. Throughout Phase I and II of this SBIR project (HL125484), we developed a panel of five critical coagulation assays on our near patient digital microfluidic instrument. The assays include antigen assays for von Willebrand factor (VWF) and Protein S, plus activity assays for Antithrombin III (ATIII), Protein C, and Factor VIII. All 5 assays are collectively run from one drop of whole blood sample (< 50 L). Additional Phase IIB funding will allow us to develop complementary molecular assays for Factor V Leiden and Prothrombin (Factor II) genetic mutations, which would be automated on the same assay platform. The addition of these molecular assays will provide a more comprehensive evaluation of coagulation risk and will improve the commercial potential of our final product. We will also perform CLSI guided analytical and clinical validations of the complete panel of coagulation tests and prepare regulatory applications for 510(k) FDA clearance. There is currently no single, small volume testing platform on the market that can support rapid testing of antigen, activity, and molecular analytes to provide a comprehensive assessment of hypercoagulability risk. Our testing platform and assays have been purposefully designed to meet the technical requirements for rapid, low complexity testing from small sample volumes and address a critical unmet need in pediatric coagulation management. Our approach is strengthened by collaborations with our clinical partner at Boston Children’s Hospital, the high potential clinical impact of the tests, the novel application of digital microfluidic technology for whole blood testing near the patient, and by thorough market analysis and other strategic marketing that has been initiated for the product through other sources of funding. The digital microfluidic assays will be suitable for use near the patient with minimal training requirements. Lead intended users of our tests will be nurses or laboratory technicians in pediatric care settings, but we anticipate that the system can be readily expanded to other markets, such as adult cardiac patients and rural or limited resource hospital settings that are not currently capable of performing the full coagulation assay panel.