A Flow Through Cell Counting Assay

流式细胞计数测定

• 批准号: 8701344
• 负责人: Simon Bystryak
• 金额: $ 46.4万
• 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 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8701344
• 关键词: Acquired Immunodeficiency Syndrome; Address; Adult; Affect; Africa South of the Sahara; Anti-Retroviral Agents; Belief; Biological Assay; Blood; Blood Glucose; Businesses; CD4 Lymphocyte Count; CD4 Positive T Lymphocytes; Caring; Cell Count; Childhood; Clinical; Cost Analysis; Country; Data Reporting; Developing Countries; Development; Devices; Disease Management; Educational Status; Epidemic; Equipment; Flow Cytometry; Goals; Gold; HIV; HIV Infections; Hand; Human; Laboratories; Laboratory Technicians; Legal patent; Leukocytes; Licensing; Manuals; Measurement; Measures; Membrane; Methods; Monitor; Patients; Performance; Phase; Procedures; Process; Production; Public Health; Reading; Reagent; Reproducibility; Research; Resources; Sampling; Sensitivity and Specificity; Signal Transduction; Small Business Innovation Research Grant; Speed; Staging; Techniques; Technology; Testing; Therapeutic; Training; Validation; Whole Blood; World Health Organization; antiretroviral therapy; assay development; base; clinical practice; clinically significant; cost; design; falls; glucose monitor; innovation; instrument; instrumentation; meetings; miniaturize; new technology; novel; point of care; prognostic; prototype; public health relevance; treatment strategy

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 potentially be completed in less than 15 minutes and can be performed by laboratory technicians with low levels of training. The specific aims of Phase II are: 1) To design and assemble a pre-production ¿-prototype of the FTCA cassette and adapt a blood glucose meter for the FTCA signal measurements; 2) To explore all variables affecting the performance of the FTCA CD4+ T-cell counting method and optimize the performance of the assay; 3) To prepare lyophilized immuno-reagents for performing FTCA and examine the stability of all FTCA reagents; 4) To validate the FTCA by examining a statistically significant number of clinical samples. 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.

描述（由申请人提供）：拟议研究的长期目标是开发一种新型定量流式细胞计数方法，用于计数人类血液中的 CD4+ T 淋巴细胞，以对 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，并且用于 HIV 分期和治疗决策 目前用于 CD4 细胞定量的黄金标准是流量。然而，流式细胞仪（包括最近开发的小型单一用途 CD4 细胞计数设备）价格昂贵，而且在资源有限的国家难以获得。此外，它们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。基于细胞计数的 CD4 细胞计数超出了许多 HIV 感染者的能力范围，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。这个上下文成本较低、快速且易于执行的方法，即 为了摆脱传统测定的限制，建议开发用于测定 CD4+ T 细胞的方法，流通细胞计数测定 (FTCA) 基于特殊膜和专有流通盒的使用。测量细胞计数状态。该测试可能会在 15 分钟内完成，并且可由受过较低培训的实验室技术人员执行。第二阶段的具体目标是：1) 设计和组装预生产 ¿ - FTCA 盒原型并采用血糖仪进行 FTCA 信号测量； 2) 探索影响 FTCA CD4+ T 细胞计数方法性能的所有变量并优化检测性能 3) 准备冻干免疫； -用于执行 FTCA 的试剂并检查所有 FTCA 试剂的稳定性；4) 通过检查统计上显着数量的临床样本来验证 FTCA 所提出的技术的优点。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。