Enhancing Pediatric Diagnosis of Tuberculosis with FLOW Technology

摘要 虽然正确的诊断是有效治疗结核病的关键，但在资源匮乏的情况下，结核病检测存在特定的障碍 环境，特别是针对儿童的。虽然有许多结核病诊断工具可用，但它们都患有严重的 缺点(困难、不准确、昂贵)，限制了它们在低资源环境中的广泛实施 以及它们在儿童中的可行性。一种分枝杆菌细胞壁糖脂，脂阿拉伯甘露聚糖(LAM)，作为诊断 病原体生物标记物已经被广泛证明，并且一点护理(POC)LAM试验(ALERE确定™ TB LAM)目前在市场上。Alere的技术将LAM检测整合到基于尿液的侧向流动中 虽然LFA易于使用，但其较差的诊断敏感性限制了其对极高血吸虫病患者的实用性 LAM浓度(例如，晚期HIV疾病患者和CD4计数&100)。这一限制是 在儿童中尤其有问题，因为他们的少杆菌结核病的发病率较高(当LAM浓度 可能更低)。 在这项研究中，我们将弥合儿童对结核病诊断的未得到满足的需求和LAM使用之间的差距 作为一种很有前途的解决方案，它应用了一种名为“Flow”的新兴技术，以增强 林丽发。虽然Alere LFA只从几滴尿液中测量LAM，但Flow从 将几毫升的尿液倒入~100微米的L中，然后用LFA进行测定。重要的是，流量集中是 完全被动，除了基于LFA的分析之外，只需要很少或不需要额外的步骤，因此使其成为一个简单的 以及用户友好的LFA“前端”，特别是在结核病最突出的低资源环境中。AS 在南非进行的一项小型、仅限成人参与的人类(FIH)研究的原则证明(由本条例草案资助 和梅琳达·盖茨基金会，见支持信)，在Alere和(我们的新产品)上运行流动浓缩尿液 已开发的Salus LFA导致临床敏感性加倍。 在这份FastTrack SBIR建议中，我们将优化和调整Flow，用于诊断儿科患者的结核病 通过：目标1，优化流动技术，以最大限度地提高结核病儿童的分析灵敏度；在目标1 2，在我们的合作临床研究伙伴进行的一项仅针对儿科的小型先导性研究中评估临床敏感性 在南非的网站；在目标3中，根据需要实施必要的改变和/或改进并制定 为我们的设备改进了收集到答案的工作流程，包括进一步开发Salus LFA。 此外，我们建议开发一种面向非常年轻的(例如0-4岁)和 病情严重，这将插入我们的流程工作流，并允许高效地收集和评估样本 患者结核病状态；在目标4中，使用我们的流动结核病检测返回南非，以获得适当的电源 研究，其结果将为我们的商业化道路(目标5)奠定基础。重要的是，我们 已确定要完成的合作伙伴(雅培实验室、直觉生物科学公司，请参阅支持函) 如果从拟议的工作中获得的数据是有希望的，那么商业化进程就会开始。

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.