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

点击翻译按钮获取中文摘要

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.