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

项目总结/摘要 尿液是一种有吸引力的即时诊断生物标本，因为它可以大量收集 采用非侵入性手术。基于尿的侧流测定（LFA）是适合于在尿液中进行检测的低成本装置。 即时检测，特别是在资源匮乏的环境中。然而，许多尿基LFA表现出敏感性， 由于尿中存在的诊断生物标志物的浓度低，其水平远低于诊断实用性。这 LFA用于结核病（TB）诊断的情况是如此，并且是快速TB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检测，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 可访问性，分别。这些测试还需要痰样本，这带来了额外的困难， 采集过程中医护人员的暴露风险。 为了解决这个问题，我们开发了一种渗透处理器， 用于LFA的生物标志物。去除了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 在早期原型中显示出20 mL样品的近100倍浓度。通过利用 渗透，最近的原型开发的公司显示承诺，在显着减少处理时间 在30分钟内。由于其简单性和灵活性，该设备展示了很大的潜力， 与现有的LFA结合，能够高灵敏度检测尿液中稀释的目标分析物，同时仍保留 诊断时间适合于护理点诊断。该项目旨在验证分子浓度 定量与质谱在较新的原型，执行加标和回收率测试与模拟 尿液和基于实验室的LAM，然后用TB阳性临床样品和LFA进行定性检测。 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 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，并计划扩大装置生产规模，以供商业销售， 第二阶段。

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.