Proof-of-Concept for Card-Based CD4 Cell Counting

描述（由申请人提供）：这项拟议研究的长期目标是开发一种简单，廉价的即时护理设备，该设备将从全血中纯化CD4+ T细胞并执行绝对CD4+ T细胞计数。这将通过调整、组合和测试一个基于芯片的原型流动粒子计数器与一个单独的被动微流控细胞分离和纯化技术来完成。为了实现这一目标，第一阶段将研究四个目标，重点是建立一个概念验证系统，使用这些技术来计数CD4+ T细胞。目的一将确定使用能够从全血中纯化CD4+ T细胞的CD4被动微流控“预备卡”的可行性。目标二将确定在一次性平台中使用微加工芯片对隔离细胞进行计数的可行性。目的三将确定使用简单的电子接口来可靠地计数CD4+T细胞的可行性。目标四将使用市售的正常血液标本测试和验证单个CD4计数成分。在完成这四个目标后，第二阶段将专注于将微流控分离“准备卡”与微加工计数器芯片集成，生产单组分、一次性使用的CD4计数器盒。然后，这个新系统将进行进一步的验证和优化，为针对CD4分析的现有行业黄金标准进行现场试验做准备。当在拟议的使用场景中进行充分验证和评估时，所得到的设备将是简单、可靠和独特的，因为它可以提供完全定量的CD4+ T细胞分析，仪器成本不到300美元。这几乎比现有流式细胞仪的成本低两个数量级。试剂和一次性成本也将低于任何目前可用的设备。该工具将立即应用于各级公共卫生保健系统，特别是在资源贫乏地区和发展中国家，用于监测人类免疫缺陷病毒（HIV）感染者，制定鉴别诊断，并就抗逆转录病毒药物治疗方案作出治疗决定，包括高效抗逆转录病毒疗法（HAART）方法。

DESCRIPTION (provided by applicant): The long-term objective of this proposed research is to develop a simple, inexpensive point-of-care device that will purify CD4+ T cells from whole blood and perform absolute CD4+ T cell counts. This will be accomplished by adapting, combining, and testing a prototype chip-based flow-through particle counter with a separate passive microfluidic cell separation and purification technique. Toward this objective, Phase I will investigate four aims focused on establishing a proof-of-concept system using these technologies to count CD4+ T cells. Aim One will determine the feasibility of using a CD4 passive microfluidic "prep card" capable of purifying CD4+ T cells from whole blood. Aim Two will determine the feasibility of using microfabricated chips to count the isolated ceils in a disposable platform. Aim Three will determine the feasibility of using a simple electronic interface to reliably count the CD4+T cells. Aim Four will test and validate the individual CD4 counting components using commercially available normal blood specimens. Upon completion of these four aims, Phase II will focus on integrating the microfluidic separation "prep card" with the microfabricated counter chips to produce a single component, one-time use CD4 counter cartridge. This new system will then undergo further validation and optimization in preparation for field trials against existing industry gold standards for CD4 analysis. The resulting device, when fully validated and evaluated in the proposed scenarios of use, will be simple, robust, and unique in that it provides a fully quantitative CD4+ T cell analysis with an instrument costing less than $300. This is almost two orders of magnitude below the cost of established flow cytometry devices. Reagent and disposable costs will also be lower than that of any currently available device. The tool will find immediate application in all levels of the public health care system, particularly in resource-poor areas and developing countries, for monitoring persons with human immunodeficiency virus (HIV), formulating differential diagnoses, and making therapeutic decisions regarding anti-retroviral drug treatment regimes, including the highly active anti-retroviral therapy (HAART) approach.