Enabling standardized point-of-care tuberculosis diagnostics via rapid osmotic concentration

通过快速渗透浓度实现标准化床旁结核病诊断

• 批准号: 10697134
• 负责人: James Lai
• 金额: $ 27.26万
• 依托单位: TRULY TECHNOLOGIES LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697134
• 关键词: 2019-nCoV; Address; Affect; Antigens; Area; Award; Biological Assay; Biological Markers; Buffers; Businesses; Centrifugation; Cessation of life; Clinical; Collaborations; Collection; Data; Detection; Devices; Diagnosis; Diagnostic; Diagnostic Procedure; Diffusion; Disease; Enzyme-Linked Immunosorbent Assay; Equation; Equipment; Excision; Exhibits; Grant; Healthcare; Human; Human Chorionic Gonadotropin; Immunoassay; Individual; Industry; Interview; Kinetics; Laboratories; Letters; Mainstreaming; Marketing; Mass Spectrum Analysis; Measures; Membrane; Methods; Microscopy; Modeling; Monitor; Noise; Nucleic Acid Amplification Tests; Nucleocapsid Proteins; Osmosis; Performance; Persons; Phase; Polymers; Power Sources; Preparation; Procedures; Process; Production; Proteins; Recovery; Research; Resource-limited setting; Risk; Sampling; Scientist; Signal Transduction; Small Business Innovation Research Grant; Solubility; South Africa; Special Equipment; Specimen; Speed; Sputum; Standardization; Surface; Temperature; Testing; Therapeutic; Time; Training; Tuberculosis; Ultracentrifugation; Ultrafiltration; Urea; Urea Nitrogen; Urine; With laterality; commercialization; cost; design; detection limit; diagnostic biomarker; diagnostic value; experience; flexibility; improved; infection risk; innovation; lateral flow assay; lipoarabinomannan; manufacturing scale-up; mycobacterial; point of care; point of care testing; point-of-care diagnostics; prevent; prototype; scale up; small molecule; success; tuberculosis diagnostics; tuberculosis treatment; urinary

PROJECT SUMMARY/ABSTRACT Urine is an attractive biospecimen for point-of-care diagnostics because it can be collected in large quantities with noninvasive procedures. Urine-based lateral flow assays (LFAs) are low-cost devices suitable for point-of-care testing, particularly in low-resource settings. However, many urine-based LFAs exhibit sensitivity levels well below diagnostic utility due to the low concentration of diagnostic biomarkers present in urine. This is the case with LFAs for tuberculosis (TB) diagnostics and is a major barrier to rapid TB testing and treatment in endemic areas. Mainstream clinical and laboratory tests for diagnosing active TB, including bacterial culture, sputum smear microscopy and nucleic acid amplification tests, present limitations in speed, sensitivity, accessibility, respectively. These tests also require sputum samples that present additional difficulty and exposure risks for healthcare staff during the collection procedure. To address this, we developed an osmotic processor that statically and spontaneously concentrates urinary biomarkers for use in LFAs. Urea and small molecules that can interfere with downstream assays are removed in the concentration process. Using human chorionic gonadotropin (hCG) protein as a model analyte, we showed near 100-fold concentration of a 20 mL sample in an early prototype. By exploiting the principles of osmosis, recent prototypes developed by the company show promise in significantly reducing processing time to within 30 minutes. With its simplicity and flexibility, the device demonstrates a great potential to be interfaced with existing LFAs to enable highly sensitive detection of dilute target analytes in urine, while still retaining a diagnostic time suitable for point-of-care diagnostics. The project aims to validate molecule concentration quantitatively with mass spectrometry in newer prototypes, perform spike-and-recovery tests with simulated urine and lab-based LAM, and then test qualitatively with TB-positive clinical samples and LFAs. If awarded the Phase I SBIR grant, the team will hire another full time research scientist, register our device as a General Controls Device through the FDA, continue business collaborations with companies developing LFAs, research other applications of the device to process other biomarkers for diagnostic, disease monitoring and therapeutic purposes, and plan scale-up of device production for commercial distribution in preparation of Phase II.

项目概要/摘要 尿液对于即时诊断来说是一种有吸引力的生物样本，因为它可以大量收集 通过非侵入性手术。基于尿液的侧流检测 (LFA) 是一种低成本设备，适用于 即时检测，特别是在资源匮乏的环境中。然而，许多基于尿液的 LFA 表现出敏感性 由于尿液中诊断生物标志物浓度低，其水平远低于诊断效用。这 用于结核病 (TB) 诊断的 LFA 就是这种情况，并且是快速结核病检测和治疗的主要障碍 在流行地区。用于诊断活动性结核病的主流临床和实验室测试，包括细菌培养、 痰涂片显微镜检查和核酸扩增测试在速度、灵敏度、 分别是可访问性。这些测试还需要痰样本，这会带来额外的困难和 医护人员在收集过程中存在暴露风险。 为了解决这个问题，我们开发了一种渗透处理器，可以静态、自发地浓缩尿液 用于 LFA 的生物标志物。去除可能干扰下游检测的尿素和小分子 在浓缩过程中。使用人绒毛膜促性腺激素 (hCG) 蛋白作为模型分析物，我们 在早期原型中，20 mL 样品的浓度接近 100 倍。通过利用以下原则 渗透，该公司最近开发的原型有望显着缩短处理时间 至30分钟内。凭借其简单性和灵活性，该设备展现出巨大的接口潜力 与现有的 LFA 结合使用，可以高度灵敏地检测尿液中的稀释目标分析物，同时仍保留 适合即时诊断的诊断时间。该项目旨在验证分子浓度 在较新的原型中使用质谱定量分析，使用模拟进行尖峰和恢复测试 尿液和基于实验室的 LAM，然后使用结核病阳性临床样本和 LFA 进行定性测试。 如果获得第一阶段 SBIR 资助，该团队将聘请另一位全职研究科学家，将我们的设备注册为 通过 FDA 的通用控制设备，继续与开发公司的业务合作 LFA，研究该设备的其他应用，以处理用于诊断、疾病监测的其他生物标志物 和治疗目的，并计划扩大设备生产规模以进行商业分销，以准备 第二阶段。