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、廉价的即时检测设备，该设备将从全血中纯化CD4+ T细胞并进行绝对CD4+ T细胞计数。这将通过调整，组合和测试原型芯片为基础的流通颗粒计数器与单独的被动微流控细胞分离和纯化技术来实现。为了实现这一目标，第一阶段将调查四个目标，重点是建立一个使用这些技术来计数CD 4 + T细胞的概念验证系统。目的探讨一种能够从全血中纯化CD4+ T细胞的CD4被动微流控“制备卡”的可行性。目的二是确定在一次性平台中使用微加工芯片对分离细胞进行计数的可行性。目的三将确定使用一个简单的电子接口，可靠地计数的CD4+T细胞的可行性。目的四是利用市售的正常人血液标本对单个CD4计数组件进行检测和验证。在完成这四个目标后，第二阶段将专注于将微流控分离“制备卡”与微制造计数器芯片集成，以生产单组分、一次性使用的CD4计数器盒。然后，这一新系统将进行进一步验证和优化，为对照现有的CD4分析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，所产生的设备将是简单，强大和独特的，因为它提供了一个完全定量的CD4+ T细胞分析，仪器成本低于300美元。这几乎比已建立的流式细胞术装置的成本低两个数量级。试剂和一次性用品的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保健系统，特别是在资源贫乏地区和发展中国家，以监测感染人体免疫缺陷病毒（艾滋病毒）的人，作出鉴别诊断，并就抗逆转录病毒药物治疗制度，包括高效抗逆转录病毒疗法（HAART）办法作出治疗决定。

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.