Enhancing Pediatric Diagnosis of Tuberculosis with FLOW Technology

利用 FLOW 技术增强儿科结核病诊断

• 批准号: 10548952
• 负责人: Brianna Mullins
• 金额: $ 96.29万
• 依托单位: SALUS DISCOVERY, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-10 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10548952
• 关键词: AIDS/HIV problem; Adult; Age; Back; Benchmarking; Biological Assay; Biological Markers; Biological Sciences; CD4 Lymphocyte Count; Cause of Death; Cell Wall; Cessation of life; Child; Childhood; Clinical; Clinical Microbiology; Clinical Sensitivity; Collection; Data; Detection; Development; Device or Instrument Development; Devices; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic Sensitivity; Disease; Effectiveness; Emerging Technologies; Enrollment; Evaluation; Feedback; Foundations; Freezing; Funding; Generations; Glycolipids; Goals; HIV; Immunocompromised Host; Incidence; Intuition; Laboratories; Letters; Measures; Patients; Performance; Phase; Pilot Projects; Population; Preparation; Process; Reagent; Recording of previous events; Resource-limited setting; Route; Running; Sampling; Sensitivity and Specificity; Small Business Innovation Research Grant; Source; South Africa; Sputum; Stomach; Technology; Testing; Tuberculosis; Tuberculosis diagnosis; Urine; Work; base; clinical research site; commercialization; cost; design; diagnostic assay; diagnostic biomarker; diagnostic tool; first-in-human; human study; improved; interest; lateral flow assay; lipoarabinomannan; lymph node biopsy; mycobacterial; new technology; participant enrollment; pathogen; pediatric patients; point of care; research clinical testing; sample collection; success; tuberculosis diagnostics; tuberculosis treatment; user-friendly

ABSTRACT While proper diagnosis is key to effective TB treatment, there exist specific barriers to TB testing in low resource settings, especially for children. While a number of TB diagnostic tools are available, they each suffer from critical drawbacks (difficult, inaccurate, expensive) that limit their widespread implementation in low resource settings and their feasibility in children. A mycobacterial cell wall glycolipid, lipoarabinomannan (LAM), as a diagnostic pathogen biomarker has been widely demonstrated and one point-of-care (POC) LAM assay (Alere Determine™ TB LAM) is currently on the market. Alere’s technology incorporates LAM detection in a urine-based lateral flow assay (LFA) and while easy to use, its poor diagnostic sensitivity limits its utility to patients with extremely high LAM concentrations (e.g., patients with advanced HIV disease and CD4 counts <100). This limitation is particularly problematic in children, given their higher incidence of paucibacillary TB (when LAM concentration is likely even lower). In this study we will bridge the gap between the unmet need for TB diagnostics in children and the use of LAM as a promising solution by applying an emerging technology, termed “FLOW”, to enhance the sensitivity of the LAM LFA. While the Alere LFA measures LAM from only a few droplets of urine, FLOW concentrates LAM from several mL of urine into ~100 µL, which is then assayed by the LFA. Importantly, FLOW concentration is completely passive, requiring little or no additional steps beyond LFA-based analysis, thus making it a simple and user friendly “front end” for LFAs, especially within low resource settings where TB is most prominent. As proof of principle, in a small, adult only, first-in-human (FIH) study performed in South Africa (funded by the Bill and Melinda Gates Foundation, see letter of support), running FLOW-concentrated urine on Alere and (our newly developed) Salus LFAs resulted in doubling of clinical sensitivity. In this FastTrack SBIR proposal we will optimize and adapt FLOW for the diagnosis of TB in pediatric patients by: In Aim 1, optimizing the FLOW technology to maximize analytical sensitivity among children with TB; In Aim 2, assessing clinical sensitivity in a small, pediatric only, pilot study performed at our partner clinical research site in South Africa; In Aim 3, implementing necessary changes and/or improvements as needed and develop an improved collection-to-answer workflow for our device, including furthering the development the Salus LFA. Additionally, we propose to develop a “wearable” adaptor, geared toward the very young (e.g. ages 0-4) and very sick, that will plug into our FLOW workflow and allow for efficient sample collection and assessment of patient TB status; and in Aim 4 employing our FLOW TB assay back to South Africa for a properly powered study, the results of which will provide a foundation for our route to commercialization (Aim 5). Importantly, we have already identified partners (Abbott Laboratories, Intuitive Biosciences, see letters of support) to complete the commercialization process if the data from the proposed work is promising.

抽象的 虽然正确诊断是有效结核病治疗的关键，但在资源匮乏的情况下进行结核病检测存在特定障碍 尽管有许多结核病诊断工具可供使用，但每种工具都面临着严重的问题。 限制其在资源匮乏环境中广泛实施的缺点（困难、不准确、昂贵） 及其在儿童中的可行性。 病原体生物标志物已得到广泛证明，并且一种即时护理 (POC) LAM 检测（Alere Define™ TB LAM）目前已上市，Alere 的技术将 LAM 检测纳入基于尿液的侧流检测中。 测定（LFA），虽然易于使用，但其较差的诊断灵敏度限制了其对病情极高的患者的实用性。 LAM 浓度（例如，患有晚期 HIV 疾病且 CD4 计数 <100 的患者）。 鉴于儿童的少杆菌结核发病率较高（当 LAM 浓度 可能甚至更低）。 在这项研究中，我们将弥合儿童结核病诊断的未满足需求与 LAM 的使用之间的差距 作为一种有前途的解决方案，通过应用一种名为“FLOW”的新兴技术来提高 LAM LFA。Alere LFA 仅从几滴尿液中测量 LAM，而 FLOW 则从尿液中浓缩 LAM。 将几毫升尿液注入约 100 µL，然后通过 LFA 进行测定。重要的是，FLOW 浓度为 完全被动，除了基于 LFA 的分析之外，几乎不需要或不需要额外的步骤，从而使其变得简单 以及用户友好的 LFA“前端”，特别是在 TB 最突出的低资源环境中。 原理证明，在南非进行的一项小型、仅限成人、首次人体 (FIH) 研究中（由该法案资助） 和梅琳达·盖茨基金会，请参阅支持信），在 Alere 上运行 FLOW 浓缩尿液，以及（我们新推出的 Salus LFA 使临床敏感性加倍。 在此 FastTrack SBIR 提案中，我们将优化和调整 FLOW 以诊断儿科患者的结核病 通过： 在目标 1 中，优化 FLOW 技术以最大限度地提高结核病儿童的分析灵敏度； 2，在我们的合作伙伴临床研究中进行的一项小型儿科试点研究中评估临床敏感性 在目标 3 中，根据需要实施必要的变更和/或改进并制定 我们的设备改进了从收集到答案的工作流程，包括进一步开发 Salus LFA。 此外，我们建议开发一款“可穿戴”适配器，面向幼儿（例如 0-4 岁）和 非常恶心，这将插入我们的 FLOW 工作流程，并允许有效的样本收集和评估 患者结核病状况；并在目标 4 中使用我们的 FLOW TB 检测返回南非进行适当供电 研究，其结果将为我们的商业化之路奠定基础（目标 5）。 已经确定合作伙伴（Abbott Laboratories、Intuitive Biosciences，请参阅支持信）来完成 如果拟议工作的数据有希望，则可以进行商业化进程。