Enabling Point of Care Screening for Sepsis Using Rapid Single Cell Analysis

项目摘要 脓毒症是一种对局部细菌或真菌感染的不受控制的全身炎症免疫反应，是 造成的死亡人数超过前列腺癌、乳腺癌和艾滋病的总和 40%的ICU费用，是美国最昂贵的住院费用(约为美国每年医疗保健的200亿美元 2011年的支出)。虽然已经证明，开始积极治疗的时间是至关重要的 改善结果(目前死亡率为30%-50%)和降低成本(&gt；22,000美元/病例)，紧急情况 内科医生在急诊中缺乏令人信服的诊断工具来检测脓毒症 科室(ED)，80%的败血症病例来源于医院。CytoVale旨在改善脓毒症 治疗模式，通过提供一个平台，在最初出现时检测异常的全身炎症。我们的 诊断平台将提供经济高效、强大和快速的样本到决策分析(&lt；10分钟)，在 表明全身炎症失控的激活的白细胞在一种 无标签的方式。为了影响护理质量，仪器必须位于护理点 (POC)，在那里它可以提供快速结果以影响患者护理。目前，由于手动操作，这是不可行的 为化验准备样品所需的操作(例如，离心法、移液法)以及缺乏内标物。 样品校准。在这里，我们建议利用我们的第一阶段自动化进展的成功 通过现在开发结合样品的完全集成的模块来制备样品的过程 制备和变形性细胞仪微流控成单一的试剂盒和仪器。这将允许 在ED POC通过指刺对疾病过程中的早期患者进行识别和分类 血。额外的成本和患者舒适度得益于能够跟踪病程和 设计针对患者反应的治疗方法；节省住院天数和相关成本。 Cytovale还建议开发和集成一种可变形的样品控制。机械标准粒子 将在每次样品运行中用作内部控制，以确保仪器读数和 诊断准确率。这不仅有助于验证实验的可靠性和一致性，而且还将提供 这是细胞生物物理学不断发展的领域中亟需的校准粒子。综合起来，这些目标将 在POC扩展强大的基于细胞的分析的可访问性。

Project Summary Sepsis, an uncontrolled systemic inflammatory immune response to local infection by bacteria or fungi, is responsible for more deaths than prostate cancer, breast cancer, and AIDS combined, accounts for more than 40% of ICU costs, and is the most expensive inpatient condition in the U.S. (~$20B in annual U.S. healthcare expenditures in 2011). While it has been demonstrated that the time to initiating aggressive treatment is critical to improving outcomes (currently 30-50% mortality) and decreasing costs (>$22,000/case), emergency medicine physicians suffer from a lack of compelling diagnostic tools for detection of sepsis in the emergency department (ED), where >80% of sepsis cases originate in the hospital. CytoVale aims to improve the sepsis treatment paradigm by offering a platform to detect abnormal systemic inflammation at initial presentation. Our diagnostic platform will offer a cost-effective, robust, and rapid sample-to-decision assay (< 10 minutes), in which activated white blood cells that are indicative of uncontrolled systemic inflammation are interrogated in a label-free manner. In order to impact quality of care, the instrument must be situated at the point-of-care (POC), where it can provide rapid results to impact patient care. Currently, this is not feasible, due to manual operations required to prepare samples for the assay (e.g., centrifugation, pipetting) and the lack of an in- sample calibration. Here, we propose to capitalize on the success of our Phase 1 progress of automating the process of sample preparation by now developing a fully integrated module that combines the sample preparation and deformability cytometry microfluidics into a single cartridge and instrument. This will allow for the identification and triaging of patients earlier in their disease process at the ED POC from a finger-prick of blood. Additional cost and patient comfort benefits arise from the ability to track the course of disease and design treatments specific to patient response; saving days spent in the hospital and associated costs. Cytovale also proposes to develop and integrate a deformable sample control. Mechanical standard particles will be used as internal controls within each sample run, to ensure fidelity of the instrument readings and diagnostic accuracy. Not only will this help verify reliability and consistency of experiments, but will also supply a much needed calibration particle to the growing field of cellular biophysics. Combined, these aims will expand accessibility of powerful cell-based assays at the POC.