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（由申请方提供）：拟定研究的长期目标是开发一种用于人血液中CD4+ T淋巴细胞计数的新型定量流通细胞计数方法。需要CD4细胞计数来分期和监测HIV感染患者。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，并用于HIV分期和治疗决策。目前用于定量CD4细胞的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细胞计数装置，价格昂贵，在资源有限的国家难以获得。此外，它们还需要训练有素的实验室技术人员来处理和加工样品，分析和报告数据。由于基于流式细胞术的CD4细胞计数超出了如此多的艾滋病毒感染者的能力范围，世界卫生组织（WHO）敦促开发用于资源有限环境的简单的即时CD4细胞计数测定。正是在这种情况下，较便宜，快速和易于执行的方法，这是免费的传统测定的局限性，用于确定CD4+ T细胞的建议被开发。 拟议的方法，即流通细胞计数测定（FTCA），是基于使用特殊膜和专有的流通盒来测量细胞计数状态。该测试可以在不到15分钟的时间内完成，并且可以由受过低水平培训的实验室技术人员进行。第I阶段的具体目标包括设计和测试FTCA试剂卡的概念验证原型，使用几种市售膜和与不同来源标记物结合的各种抗CD4抗体检查FTCA的性能，以及优化用于进行FTCA的试剂浓度和测定条件。 所提出的技术的优点，如分析的速度，方便，和低成本证实了我们的信念，FTCA将在临床实践中广泛使用。 公共卫生相关性：拟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。这些研究的成功完成将有利于公众健康，并提供大量商业机会所需的所有技术，以许可该技术并制造商业产品。