Ultra-Stable Test Strip System for Multiplexed Detection of Enteric Pathogens

描述（由申请人提供）：腹泻是全球死亡的主要原因，特别是在获得医疗资源有限的婴儿和幼儿中。虽然准确和及时地诊断致病传染病将提高存活率并减轻流行病的传播，但目前在不可控环境中运输和储存敏感诊断的挑战在经济上或后勤上都是不可实现的。为了满足这一需求，智能光学系统（IOS）和合作伙伴通用稳定技术（UST）提出创建一个超稳定的一步免疫分析平台，可以在单个样品中以高灵敏度和特异性识别多种肠道病原体。这种快速和廉价的即时检测将准确识别一种或多种病原体，使卫生工作者能够启动最适当的治疗，而不会产生产生抗生素耐药病原体的不必要风险。为了鼓励世界范围内的采用和使用，该测试还将易于使用，非常稳定，对现场部署具有长期功效，敏感和微生物特异性，并且价格低廉。现有的免疫诊断在该领域是不充分和不切实际的，因为它们通常需要昂贵和耗电的制冷来在不可控的环境条件下保持功效。此外，许多免疫测定的有效执行也需要多步骤方案的培训和经验，以产生有意义的结果。不幸的是，在教育、资源和社会基础设施匮乏的发展中地区，即使是进行免疫测定的最基本要求，即电力、清洁水和培训，也非常昂贵或根本不可能。在第一阶段，IOS将开发一种横向流动免疫测定（LFIA）测试条格式，能够在单个样品中检测多种感染性病原体，假阳性或假阴性发生率低。UST的专利技术可以保存试纸，即使在低于50℃的温度下延长储存两年，也能保持充分的活性。我们将建立一个简单的一步式程序，这样受过最低限度培训的工作人员即使使用原始粪便样本也能得出准确一致的结果。使用肉眼可检测的高荧光量子点（QD）探针，该敏感系统将在15分钟内产生样品中病原体的定性结果。在第二阶段，IOS将整合更多的其他粪便肠道病原体的免疫分析到相同的试纸格式，以产生一个全面的病原体筛选。对于在低病原体浓度下极具传染性的传染病，我们将建立一个小型，廉价，便携式和坚固耐用的试纸阅读器，以增强定量检测。在第二阶段，IOS还将准备材料，包括在FDA批准的申请中，以便原型系统可以进行临床评估。公共卫生相关性：每年，腹泻导致数百万人死亡，特别是五岁以下儿童。许多病原体，包括细菌、寄生虫、病毒和真菌，可能在短短72小时内引起致命的腹泻。在世界上许多经济、气候和/或地理上具有挑战性的地区，由于其蛋白质试剂的脆弱性和进行检测的技术难度，无法获得敏感的免疫测定诊断。使用简单且超稳定的试纸系统可以准确区分单个样品中的几种潜在病原体，这将促进适当的医疗干预并提供确定的治疗选择。多路复用格式还将消除多次测试的需要，降低单位成本，并扩大获得这些关键工具的途径，以防止不必要的死亡。

DESCRIPTION (provided by applicant): Diarrhea is a leading cause of death globally, especially among infants and young children with limited access to medical resources. Although accurate and timely diagnosis of the causative infectious diseases would increase survival rates and mitigate epidemic spread, the current challenges of transporting and storing sensitive diagnostics in uncontrollable environments are not economically or logistically feasible. To meet this need, Intelligent Optical Systems (IOS) and collaborator, Universal Stabilization Technologies (UST), propose to create an ultra-stable, one-step immunoassay platform that can identify multiple enteric pathogens in a single sample with high sensitivity and specificity. This rapid and inexpensive point-of-care test will accurately identify one or more pathogens so that health workers can initiate the most appropriate treatments without the unnecessary risks of creating antibiotic-resistant pathogens. To encourage worldwide adoption and use, this test will also be easy-to-use, very stable with long efficacy for field deployment, sensitive and microorganism specific, and inexpensive. Existing immunodiagnostics are inadequate and impractical in the field, because they often require expensive and power-intensive refrigeration to retain efficacy in uncontrollable environmental conditions. Moreover, the effective execution of many immunoassays also requires training and experience with multi-stepped protocols, to yield meaningful results. Unfortunately, even the most basic requirements to run immunoassays, i.e. power, clean water, and training, are prohibitively expensive or impossible in developing regions with minimal education, resources, and social infrastructure. In Phase I, IOS will develop a lateral flow immunoassay (LFIA) test strip format capable of detecting multiple infectious pathogens in a single sample with low incidence of false positives or negatives. UST's patented technology will preserve test strips so that full activity is retained even after prolong storage at temperatures <50oC over a two-year period. We will establish a simple, one-step protocol so that minimally trained workers can produce accurate and consistent results, even with raw stool samples. Using highly fluorescent quantum dot (QD) probes that are detectable with the naked eye, this sensitive system will yield qualitative results of the pathogens present in sample within 15 minutes. In Phase II, IOS will integrate more immunoassays for other fecal enteropathogens onto the same test strip format to produce a comprehensive pathogen screen. For infectious diseases that are extremely contagious at low pathogen concentrations, we will build a small, inexpensive, portable, and rugged test strip reader for enhanced quantitative detection. In Phase II, IOS will also prepare material to include in an application for FDA approval so that the prototype system can be clinically evaluated. PUBLIC HEALTH RELEVANCE: Every year, diarrhea causes the deaths of millions of people, especially children under the age of five. Numerous pathogens, including bacteria, parasites, viruses, and fungi, may cause fatal diarrhea in as little as 72 hours. In many economically, climatologically, and/or geographically challenging regions of the world, sensitive immunoassay diagnostics are unavailable because of the fragile nature of their protein reagents and the technical difficulty of performing the assays. A simple-to-use and ultra-stabilized test strip system that can accurately differentiate between several potential pathogens in a single sample would facilitate appropriate medical intervention and provide a certainty of treatment options. A multiplexed format would also eliminate the need for multiple tests, lower the unit cost, and broaden access to these critical tools, to prevent unnecessary death.