A Flow Through Cell Counting Assay

流式细胞计数测定

• 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
• 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

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）：拟议的研究的长期目标是通过细胞计数方法开发一种新的定量流，以枚举人类血液中CD4+ T淋巴细胞的枚举。需要进行CD4细胞计数来分阶段和监测HIV感染的患者。在成年人中，每微升血液的CD4+ T细胞的绝对数量具有关键的预后和治疗意义，并且用于HIV分期和治疗决策。当前用于定量CD4细胞的金标准技术是流式细胞术。但是，流式细胞仪，包括最近开发的较小的单能CD4细胞计数设备，在资源有限的国家 /地区不可接近。此外，他们要求训练有素的实验室技术人员处理和处理样本，并分析和报告数据。由于基于流式细胞仪的CD4细胞计数超出了许多受HIV感染的人的影响力，因此世界卫生组织（WHO）敦促开发为资源有限设置设计的简单护理CD4细胞计数分析。正是在这种情况下，提议开发CD4+ T细胞的较便宜，快速且易于执行的方法，该方法没有常规测定的局限性。 所提出的方法，流通电池计数测定法（FTCA）是基于使用特殊膜和专有流通盒来测量细胞计数状态的使用。该测试可以在不到15分钟的时间内完成，可以由培训水平较低的实验室技术人员进行。第一阶段的具体目的包括设计和测试FTCA盒的概念验证原型，使用几种市售膜检查FTCA的性能以及各种抗CD4抗体与不同起源和优化浓度的标签结合的各种抗CD4抗体进行FTCA的试剂和测定条件。 诸如分析的速度，便利和低成本之类的拟议技术的优势证明了我们相信FTCA将被广泛用于临床实践。 公共卫生相关性：所提出的方法的开发将允许使用CD4细胞计数测试进行分期和监测HIV感染。这些研究的成功完成将对公共卫生有益，并为有实质性的商业机会提供许可和制造商业产品所需的所有技术。