Enhancing detection and mobile monitoring of schistosomiasis with urine-based analyte pre-concentration technology

利用基于尿液的分析物预浓缩技术加强血吸虫病的检测和移动监测

• 批准号: 10697011
• 负责人: Cody Carrell
• 金额: $ 27.55万
• 依托单位: SALUS DISCOVERY, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697011
• 关键词: Academic Medical Centers; Address; Adult; Antigens; Area; Biological Assay; Blood; Buffers; Centrifugation; Cessation of life; Chronic Disease; Clinical; Clinical Research; Clinical Sensitivity; Clinical Trials; Communicable Diseases; Communities; Consumption; Data; Detection; Devices; Diagnostic; Diagnostic Sensitivity; Diagnostic Specificity; Diagnostic tests; Disease; Feces; Friends; Goals; HIV; HIV/AIDS; Human; Immunoassay; Improve Access; Individual; Infection; Infrastructure; Interruption; Laboratories; Lateral; Malaria; Maps; Measures; Methods; Microscopy; Monitor; Morbidity - disease rate; Parasitic Diseases; Patients; Performance; Persons; Pharmaceutical Preparations; Phase; Pilot Projects; Population; Prevalence; Process; Reproducibility; Research; Resources; Sampling; Schistosoma; Schistosoma mansoni; Schistosomiasis; Serum; Site; Small Business Innovation Research Grant; Snails; Specificity; Techniques; Technology; Testing; Time; Training; Treatment Efficacy; Urine; Water; World Health Organization; antigen detection; burden of illness; commercialization; cross reactivity; detection limit; detection method; disability-adjusted life years; disorder control; egg; experience; field study; first-in-human; global health; health organization; human study; improved; interest; lateral flow assay; new technology; performance tests; point of care; prototype; response; success; transmission process; treatment response; vector

ABSTRACT Schistosomiasis is a debilitating parasitic disease that infects over 230 million people and causes 280,000 deaths per year. New estimates of global disease burden rank schistosomiasis as the third most impactful infectious disease behind only HIV/AIDS and malaria. Therefore, over the past decade, global health organizations have renewed interest in schistosomiasis control and elimination. Efforts to reduce schistosomiasis-associated morbidities primarily involve mass drug administration (MDA) and measures for interrupting transmission (e.g., via improved access to safe water and snail (vector) control). Critical to both disease control strategies is the ability to detect infected individuals; however, because highly endemic areas often lack basic laboratory infrastructure, it is vital that monitoring be performed in the field or at the point-of-care (POC) to address the vast numbers of sites and individuals needing testing. Current disease monitoring largely depends upon microscopy techniques to identify/quantify eggs that are sporadically shed by adult worms in stool or urine. While highly specific, microscopy methods are laborious, time-consuming, inconsistent, and often have clinical sensitivities under 5%. Alternatives to microscopy include immunoassays that detect schistosome- specific antigens, such as the circulating cathodic antigen (CCA), or the circulating anodic antigen (CAA), from human urine. A POC CCA urine assay has been commercialized and used in surveillance and mapping studies, but is not sensitive enough to detect low-intensity infections, and is only reliably produced by one of the six common Schistosoma species, which limits its use to certain regions. In contrast, CAA is produced by all Schistosoma species and, unlike microscopy, is directly correlated to total worm burden for monitoring therapy response. Therefore, a CAA immunoassay is expected to significantly enhance diagnostic monitoring of schistosomiasis, but no commercially available POC-CAA assay exists. To address this unmet need, we will integrate two complementary technologies that enable ultra-sensitive POC field-testing for schistosomiasis from urine. Our collaborator, Dr. Paul Corstjens, a globally recognized schistosomiasis expert at Leiden University Medical Center, has developed an ultrasensitive Up-Converting Phosphor Lateral Flow Assay (UCP-LFA) for CAA detection. The assay has shown tremendous promise in several studies and clinical trials but still requires time- consuming and resource-intensive sample pre-concentration to reach the sensitivity required to detect the lowest-intensity infections. In parallel, Salus Discovery has developed a new technology, termed FLOW™, that expands upon the operational concepts of LFAs by enabling pre-concentration of analytes from 20 mL of urine into 100 µL prior to detection on an LFA. Recently, Salus and Dr. Corstjens' group developed the first prototype of a device that integrates FLOW urine pre-concentration with a UCP-LFA readout (FLOW-S). The FLOW-S prototype was preliminarily evaluated with a set of 30 clinical samples where it achieved 79% sensitivity and 100% specificity, demonstrating its use as a fundamentally new, POC-friendly, ultra-sensitive CAA-based assay for schistosomiasis. In this SBIR Phase 1 proposal we will build upon our success by optimizing the FLOW-S device to achieve a detection limit of 0.1 pg/mL, enabling detection of even the lowest intensity (i.e., single-worm) infections (Aim 1), and performing a clinical study with fresh urine samples (Aim 2).

摘要 血吸虫病是一种使人衰弱的寄生虫病，每年感染2.3亿人，造成28万人死亡。 对全球疾病负担的新估计将血吸虫病列为第三大影响最大的传染病，仅次于 艾滋病毒/艾滋病和疟疾。因此，在过去的十年里，全球卫生组织重新对 控制和消灭血吸虫病。减少血吸虫病相关发病率的努力主要涉及大规模 药物管理(MDA)和阻断传播的措施(例如，通过改善获得安全水和蜗牛的机会 (向量)控制)。这两种疾病控制策略的关键是检测感染者的能力；然而，因为 高度流行的地区往往缺乏基本的实验室基础设施，因此至关重要的是在实地或在 护理点(POC)解决大量需要测试的站点和个人。 目前的疾病监测在很大程度上依赖于显微镜技术来识别/量化零星排出的鸡蛋。 由成虫在粪便或尿液中。虽然高度特异性，显微镜方法是费力、耗时、不一致的， 临床敏感度通常低于5%。显微镜的替代方法包括检测血吸虫的免疫分析- 来自人类的特定抗原，如循环阴极抗原(CCA)或循环阳极抗原(CAA) 尿液。POC CCA尿液分析已经商业化，并用于监测和地图研究，但不敏感 足以检测到低强度的感染，并且只由六种常见血吸虫物种中的一种可靠地产生， 这限制了它在某些地区的使用。相反，CAA是由所有血吸虫物种产生的，与显微镜不同，它是 与总蠕虫负担直接相关，用于监测治疗反应。因此，CAA免疫分析有望 大大加强了对血吸虫病的诊断监测，但还没有商业化的POC-CAA检测方法。 为了满足这一未得到满足的需求，我们将整合两项互补技术，以实现超灵敏的POC现场测试 用于尿液中的血吸虫病。我们的合作者保罗·科斯特延斯博士是莱顿大学全球公认的血吸虫病专家 大学医学中心，已开发出一种超灵敏的上转换磷光体横向流动分析(UCP-LFA)，用于 CAA检测。这种分析方法在几项研究和临床试验中显示出了巨大的前景，但仍需要时间- 消耗和资源密集的样品预浓缩，以达到检测最低强度所需的灵敏度 感染。与此同时，Salus Discovery开发了一种名为Flow™的新技术，该技术在 通过允许在检测前将20毫升尿液中的分析物预浓缩到100微米L中，来实现LFA的操作概念 在LFA上。最近，Salus和Corstjens博士的团队开发了第一个集成流动尿液的设备原型 用UCP-LFA读数进行预浓缩(流动-S)。对Flow-S样机进行了初步评估 30个临床样本，获得了79%的敏感性和100%的特异性，证明了它作为一种全新的、 POC友好、超灵敏的血吸虫病CAA检测方法。在此SBIR第1阶段计划中，我们将在 成功通过流程优化-S装置，实现了0.1pg/mL的检测限，使检测甚至达到了最低 加强(即单虫)感染(目标1)，并对新鲜尿样进行临床研究(目标2)。