Rapid Sample-to-Answer Diagnosis of Kaposi's Sarcoma Across Sub-Saharan Africa using KS-COMPLETE

摘要 在这项计划中，我们将开发、制造和实施撒哈拉以南非洲地区的多地点临床 KS-Complete的验证--第一个真正的医疗点样本到答案诊断系统 卡波西肉瘤(KS)。我们最近在非洲的大规模研究表明，KS可以通过 皮肤活检组织中Kaposi肉瘤疱疹病毒(KSHV)DNA的定量检测具有高敏感性和特异性。 这些努力还导致了TINI的发展-一个强大的、易于使用的、免基础设施的点- 用于KSHV DNA量化的Of-Care(PoC)技术-该技术目前正在多个站点部署 评估。这项工作还揭示了广泛采用基于皮肤活检的PoC的关键挑战 系统是从皮肤活检中提取DNA所需的时间和手动步骤-这可能长达4小时。 KS-Complete将是第一个用于皮肤穿孔活检的“直接到灯”诊断系统。类似 直接到LAMP的方法大大简化了对除固相外的其他样本矩阵的PoC诊断， 皮肤的胶原性使这成为活组织检查的一个挑战。KS-Complete将通过以下方式解决此问题 我们的“切片机”技术会自动将穿孔活检处理成更小的“微核心”，在上面我们可以 通过我们的“直接到灯”的方法，可以直接在TING中进行DNA定量。这一方法将 将结果的时间减少到60分钟左右，消除了目前所有的人工和密集的样品处理 并与LMIC中操作的成本、坚固性、基础设施和简单性要求兼容。 该系统的临床验证将通过我们在非洲建立的KS临床站点网络进行。通过 项目结束后，我们将交付12套KS-Complete系统，并进行多点临床验证。 KS是撒哈拉以南非洲最常见的男性和女性癌症之一。KS很难区分 从其他皮肤病，特别是在非洲，那里的训练有素的病理学家有限， 免疫组织化学几乎不存在。早期和更准确的诊断将提供许多 临床益处。对于患有KS的患者，它消除了难以获得、缓慢和不可靠的需求 组织病理学，并允许在更早的临床阶段进行检测，从而获得更好的临床结果。对病人来说 利用模拟试剂，快速排除KS允许及时重新确定诊断过程的方向并防止使用 有潜在毒性的化疗。我们的直接到灯诊断测试可能会产生重大影响 KS的诊断为目前诊断的多种其他病毒、分枝杆菌和真菌相关皮肤病 通过传统的病理学可以过渡到这种方法，并最终达到护理的目的。

Abstract In this proposal, we will develop, manufacture, and perform a multi-site sub-Saharan African clinical validation of KS-COMPLETE — the first true point-of-care sample-to-answer diagnostic system for Kaposi's sarcoma (KS). Our recent large-scale studies in Africa have shown that KS can be diagnosed through quantification of Kaposi's sarcoma herpesvirus (KSHV) DNA in a skin biopsy with high sensitivity and specificity. These efforts have also resulted in the development of TINY — a robust, easy-to-use, infrastructure-free, point- of-care (PoC) technology for KSHV DNA quantification — which is being currently deployed in a multi-site evaluation. The work has also revealed that the key challenge to widespread adoption of skin biopsy-based PoC systems is the time and manual steps required to extract DNA from a skin biopsy — which can be up to 4 hours. KS-COMPLETE will be the first “direct-to-LAMP” diagnostic system for skin punch biopsies. Similar direct-to-LAMP methods have greatly simplified PoC diagnostics for other sample matrices but the solid-phase, collagenous nature of skin has made this a challenge for biopsies. KS-COMPLETE will address this issue with our “SLICER” technology that will automatically process a punch biopsy into smaller “micro-cores” on which we can directly perform DNA quantification in TINY through our “direct-to-LAMP” approach. This approach will reduce the time to result to around 60 minutes, eliminate all the current manual and intensive sample processing steps, and is compatible with cost, robustness, infrastructure, and simplicity requirements for operation in LMICs. Clinical validation of the system will be done through our established network of KS clinical sites in Africa. By the end of the project, we will deliver 12 KS-Complete systems and conduct a multi-site clinical validation. KS is one of the most common cancers in men and women in sub-Saharan Africa. KS is difficult to distinguish from other skin conditions, particularly in Africa where access to trained pathologists is limited and immunohistochemistry is practically non-existent. Early-stage and more accurate diagnosis would confer many clinical benefits. For patients who have KS, it obviates the need for the difficult to obtain, slow, and unreliable histopathology and allows for detection at earlier clinical stages resulting in better clinical outcomes. For patients with mimickers, rapid exclusion of KS allows for timely re-orienting of the diagnostic process and prevents use of potentially toxic chemotherapy. Our direct-to-LAMP diagnostic test could have significant impact beyond the diagnosis of KS as multiple other viral, mycobacterial and fungal-related skin diseases currently diagnosed through traditional pathology could be transitioned to this method and ultimately the point of care.