A Flow Through Cell Counting Assay

流式细胞计数测定

基本信息

批准号：
7840786
负责人：
Simon Bystryak
金额：
$ 18.48万
依托单位：
ALLIED INNOVATIVE SYSTEMS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-03-05 至 2011-03-04
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7840786
关键词：
Acquired Immunodeficiency Syndrome Adult Africa South of the Sahara Anti-Retroviral Agents Belief Biological Assay Blood Businesses CD4 Lymphocyte Count CD4 Positive T Lymphocytes Caring Cell Count Clinical Cost Analysis Country Data Reporting Developing Countries Development Devices Educational Status Enzymes Epidemic Equipment Evaluation Eye Flow Cytometry Goals Gold HIV HIV Infections Human Infection Label Laboratories Laboratory Technicians Leukocytes Licensing Manuals Measures Membrane Methods Metric Monitor Optics Patients Performance Phase Preventive Intervention Procedures Process Public Health Reader Reading Reagent Reporting Research Research Infrastructure Resources Sampling Signal Transduction Solutions Speed Staging System Techniques Technology Testing Therapeutic Time Training Whole Blood World Health Organization antibody conjugate antiretroviral therapy base clinical practice cost design enzyme substrate falls meetings novel point of care prognostic prototype public health relevance

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to develop a novel quantitative flow through cell counting method for enumeration of CD4+ T lymphocytes in human blood. CD4 cell counts are needed to stage and monitor HIV-infected patients. In adults, the absolute number of CD4+ T cells per microlitre of blood has critical prognostic and therapeutic implications, and is used for both HIV staging and treatment decisions. The gold standard technique currently used for quantitation of CD4 cells is flow cytometry. However, flow cytometers, including smaller single-purpose CD4 cell counting devices developed recently are expensive and inaccessible in resource-limited countries. In addition, they require trained laboratory technicians to handle and process samples and to analyze and report data. Because flow cytometry-based CD4 cell counting is beyond the reach of so many HIV-infected people, the World Health Organization (WHO) has urged the development of simple point-of-care CD4 cell count assays designed for resource-limited settings. It is in this context that the less expensive, rapid and easy-to- perform method, which is free from the limitations of the conventional assays, for determination of CD4+ T cells is proposed to be developed. The proposed method, flow-through cell counting assay (FTCA) is based on the use of the special membrane and a proprietary flow-through cassette to measure cell count status. The test can be completed in less than 15 minutes and can be performed by laboratory technicians with low levels of training. The specific aims of Phase I include designing and testing a proof-of-concept prototype of the FTCA cassette, examination of the performance of the FTCA using several commercially available membranes and various anti-CD4 antibodies conjugated with labels of different origin and optimization of concentrations of the reagents and assay conditions for performing of FTCA. Advantages of the proposed technology such as the speed, convenience, and low cost of the analysis substantiates our belief that the FTCA will be widely used in clinical practice. PUBLIC HEALTH RELEVANCE: The development of the proposed method will allow using CD4 cell counting tests for staging and monitoring HIV infection. Successful completion of these studies will be beneficial for public health, and make available all of the technology needed for a substantial business opportunity to license the technology and manufacture commercial products.

描述（由申请人提供）：本研究的长期目标是开发一种新型定量流式细胞计数方法，用于计数人血液中的 CD4+ T 淋巴细胞。需要 CD4 细胞计数来对 HIV 感染患者进行分期和监测。在成人中，每微升血液中 CD4+ T 细胞的绝对数量具有重要的预后和治疗意义，可用于 HIV 分期和治疗决策。目前用于 CD4 细胞定量的黄金标准技术是流式细胞术。然而，流式细胞仪，包括最近开发的较小的单一用途 CD4 细胞计数设备，价格昂贵且在资源有限的国家难以获得。此外，他们还需要经过培训的实验室技术人员来处理和处理样品以及分析和报告数据。由于许多 HIV 感染者无法进行基于流式细胞术的 CD4 细胞计数，因此世界卫生组织 (WHO) 敦促开发专为资源有限的环境设计的简单的护理点 CD4 细胞计数测定法。正是在这种背景下，建议开发一种更便宜、快速且易于执行的方法，该方法不受传统测定法的限制，用于测定 CD4+ T 细胞。所提出的方法，流通细胞计数测定（FTCA）是基于使用特殊的膜和专有的流通盒来测量细胞计数状态。该测试可在 15 分钟内完成，并且可由培训水平较低的实验室技术人员执行。第一阶段的具体目标包括设计和测试 FTCA 盒的概念验证原型、使用几种市售膜和与不同来源标记缀合的各种抗 CD4 抗体检查 FTCA 的性能以及浓度优化执行 FTCA 的试剂和测定条件。所提出的技术的优势，例如分析的速度、便利性和低成本，证实了我们的信念，即 FTCA 将广泛应用于临床实践。公共卫生相关性：所提议方法的开发将允许使用 CD4 细胞计数测试来分期和监测 HIV 感染。成功完成这些研究将有利于公众健康，并提供获得技术许可和制造商业产品的大量商业机会所需的所有技术。