Automated Sample Preparation for Onsite Blood-Based Single- Cell Assays

用于现场血液单细胞测定的自动化样品制备

• 批准号: 8756845
• 负责人: Ajay Shah
• 金额: $ 22.5万
• 依托单位: CYTOVALE, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8756845
• 关键词: Accident and Emergency department; Accounting; Acquired Immunodeficiency Syndrome; Adoption; Antibiotics; Area Under Curve; Automation; Bacteria; Biological Assay; Biological Markers; Blood; Blood Volume; Blood specimen; Buffers; Cells; Cessation of life; Chromatin; Clinical; Clinical Research; Clinical Trials; Complex; Cytolysis; Cytometry; Data; Databases; Detection; Development; Diagnosis; Diagnostic; Disease; Drops; Electrodes; Engineering; Erythrocytes; Etiology; Event; Excision; Expenditure; Feedback; Fingers; Flow Cytometry; Focal Infection; Goals; Grant; Healthcare; Heart; Hospitals; Immune response; Infection; Inflammation; Inflammatory; Injection of therapeutic agent; Inpatients; Label; Leukocytes; Link; Liquid substance; Lung; Malignant Neoplasms; Malignant neoplasm of prostate; Manuals; Measurement; Measures; Mechanics; Methods; Metric; Microfluidic Microchips; Microfluidics; Molds; Monitor; Nuclear Envelope; Organ failure; Outcome; Output; Participant; Patient Triage; Patients; Phenotype; Physicians; Pleural effusion disorder; Population; Preparation; Procedures; Process; Property; Proteins; Protocols documentation; Quality of Care; Reagent; Research; Sampling; Science; Sepsis; Series; Small Business Innovation Research Grant; Solutions; Specificity; Speed; Staging; Suspension substance; Suspensions; System; Technology; Testing; Time; Tissues; Triage; Whole Blood; Work; base; cohort; commercialization; cost; cost effective; empowered; fungus; improved; instrument; malignant breast neoplasm; mortality; nanofabrication; neonatal sepsis; next generation; operation; point of care; public health relevance; research clinical testing; response; scale up; screening; standard of care; therapy design; time use; tool; translational medicine

DESCRIPTION (provided by applicant): 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 ~40% mortality) and decreasing costs (>$22,000/case), the field suffers from a lack of compelling early diagnostic tools. 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 (< 5 minute), in which activated white blood cells that are indicative of uncontrolled systemic inflammation are identified 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 feedback to physicians. Currently, this is not feasible, due to manual operations required to prepare samples for the assay. Primarily, this involves removal of red blood cells, debris, and large concentrations of protein, which confound CytoVale's test. Here, we propose to automate the process of sample preparation by developing an integrated module which makes use of a microfluidic technology termed Rapid Inertial Solution Exchange (RInSE). Primarily, this will enable identification and triaging of patients at earlier sepsis disease stage from a finger-prick of blood. Additional cost and patient comfort benefits arise from the ability t track the course of disease and design treatments specific to patient response, saving days spent in the hospital and associated costs. CytoVale's test identifies activated cells through automated high-speed measurements of their mechanical properties, purportedly linked to their ability to infiltrate tissue to response to infection. The first aim of the proposal will contribut to an understanding of the effects of sample preparation, specifically red blood cell lysis, on this class of emerging biomarkers. Furthermore, the microfluidic sample preparation module will be more broadly useful amongst other cell analysis methods, such as traditional flow cytometry. Combined, these aims will expand accessibility of powerful cell-based assays at the point-of-care.

描述（由申请人提供）：脓毒症是一种对细菌或真菌局部感染的不受控制的全身炎症免疫反应，其死亡人数超过前列腺癌、乳腺癌和艾滋病的总和，占ICU费用的40%以上，是美国最昂贵的住院疾病（2011年美国年度医疗保健支出约为200亿美元）。虽然已经证明，开始积极治疗的时间对于改善结果（目前约40%的死亡率）和降低成本（> 22，000美元/例）至关重要，但该领域缺乏令人信服的早期诊断工具。CytoVale旨在通过提供一个在初始表现时检测异常全身性炎症的平台来改善脓毒症治疗模式。我们的诊断平台将提供一个具有成本效益， 稳健、快速的样品-决定测定（< 5分钟），其中以无标记的方式鉴定指示不受控制的全身性炎症的活化的白色血细胞。在 为了影响护理质量，仪器必须位于护理点（POC），在那里它可以向医生提供快速反馈。目前，这是不可行的，因为需要手动操作来制备用于测定的样品。首先，这涉及去除红细胞，碎片和大浓度的蛋白质，这混淆了CytoVale的测试。在这里，我们建议通过开发一个集成的模块，利用称为快速惯性溶液交换（RInSE）的微流体技术，自动化样品制备过程。首先，这将使患者能够在早期脓毒症疾病阶段从手指刺血中识别和分类。额外的成本和患者舒适度的好处来自于跟踪疾病过程和针对患者反应设计治疗的能力，节省了在医院花费的时间和相关成本。CytoVale的测试通过自动高速测量其机械特性来识别激活的细胞，据称这些机械特性与它们渗透组织以应对感染的能力有关。该提案的第一个目的将有助于理解样本制备，特别是红细胞裂解对这类新兴生物标志物的影响。此外，微流体样品制备模块将在其他细胞分析方法中更广泛地有用，例如传统的流式细胞术。结合起来，这些目标将扩大强大的基于细胞的检测在护理点的可及性。