SBIR Phase II: Recombinant Multi-epitope Mosaic Protein Design for Urine-based Diagnosis of Leptospirosis

这一小型企业创新研究(SBIR)第二阶段项目建议开发和验证一种非侵入性、多重尿量尺，用于临床诊断由多菌感染引起的急性发热病(AFI)。目标疾病包括钩端螺旋体病、伤寒、登革热以及恶性疟和间日疟，这些疾病每年导致全球约2.29亿AFI和80万人死亡。目前的诊断工具具有侵入性，技术复杂，因此不适用于这些疾病最普遍的热带和亚热带国家，特别是在血液检测不切实际的农村和护理地点环境中。第一阶段项目成功地开发并验证了病原体特异性重组抗体试剂，该试剂在尿液中显示了诊断潜力。这一第二阶段的项目设想了两个广泛的目标：(I)重组单抗试剂的详细特征，以验证其单独和集体诊断的有效性；以及(Ii)开发和验证一种多用途试纸检测设备，用于在单一尿样中同时检测一组导致急性发热的主要原因。可交付的产品将是一种简单的诊断设备，有可能诊断恶性疟原虫、间日疟原虫和钩端螺旋体中的一种/两种，它结合了改进AFI特定诊断的需要和解决急性发烧的其他原因的需要。如果成功，该项目的更广泛影响/商业潜力将是提供一种快速诊断工具，以解决处于低资源环境中的人们面临的一个重大挑战，即如何在没有快速鉴别诊断测试的情况下管理急性发烧。虽然对每个潜在的发烧原因进行单独的测试是有益的，但在这些设置中快速选择、运行和解释多个单独的测试通常是不可行或不实用的。这种单独测试的成本影响也将是令人望而却步的。因此，迫切需要这里设想的一种多重快速设备来评估与症状相关的发热性疾病组。它将为在单一尿样中快速检测急性发热性疾病的主要原因提供医疗和后勤方面的好处。它将提供具有更广泛影响的单一答案的多疾病诊断，并通过多路分析而不是单独检测提供鉴别诊断，最适合于诊断实验室能力有限的资源有限的环境。这种多重尿试纸条被设计为一个基础平台设备，可以反复添加其他急性发烧诊断试剂，以适应特定地区。

This Small Business Innovation Research (SBIR) Phase II project proposes to develop and validate a non-invasive, multiplex urine dipstick for clinical diagnosis of Acute Febrile Illness (AFI) caused by polymicrobial infections. The target diseases include leptospirosis, typhoid fever, dengue, as well as Plasmodium falciparum and P. vivax malaria, responsible for ~229M AFI's and 800,000 deaths a year worldwide. Current diagnostic tools are invasive, technically complex, and thus not suitable in tropical and subtropical countries where these diseases are most prevalent, especially in rural and point-of-care settings where blood testing is impractical. The Phase I project successfully developed and validated pathogen-specific recombinant antibody reagents that demonstrated diagnostic potential in urine. This Phase II project envisages two broad objectives: (i) detailed characterization of the recombinant monoclonal antibody reagents to validate their individual and collective diagnostic utility; and (ii) to develop and validate a multiplex dipstick test device for concurrent detection of a set of leading causes of acute fever in a single urine sample. The deliverable will be a simple diagnostic device with the potential to diagnose either/both Plasmodium falciparum, Plasmodium vivax and Leptospira, which combines the need to improve specific diagnosis of AFI with the need to address alternative causes of acute fever.The broader impact/commercial potential of this project, if successful, will be the delivery of a rapid diagnostic tool that addresses a major challenge faced by people in low resource settings with high burdens of global health diseases, i.e. how to manage acute fever without the benefit of rapid differential diagnostic tests. Although individual tests for each potential cause of fever would be beneficial, it is often not feasible or practical to rapidly select, run, and interpret multiple individual tests in these settings. The cost implications for such individual testing also will be prohibitive. Therefore, a multiplex rapid device as envisaged here that will evaluate groups of symptomatically related febrile diseases is urgently needed. It will offer both medical and logistical benefits for rapid testing of leading causes of acute febrile illness in a single urine sample. It will deliver a single answer multi-disease diagnostic with broader impact, and offer differential diagnosis through a multiplexed assay rather than individual testing, most suitable in resource limited settings where diagnostic laboratory capacities are limited. This multiplex urine dipstick test is designed to be a foundational platform device to which other diagnostic reagents for acute fever can be iteratively added to suit a particular region.