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

描述（由申请人提供）：腹泻是全球主要的死亡原因，尤其是在获得医疗资源有限的婴幼儿中。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，但目前在不可控的环境中运输和储存敏感诊断试剂的挑战在经济上或后勤上都不可行。为了满足这一需求，智能光学系统公司（IOS）和合作者通用稳定技术公司（UST）提出创建一种超稳定的一步免疫分析平台，该平台可以以高灵敏度和特异性在单个样本中识别多种肠道病原体。这种快速、廉价的即时检测将准确识别一种或多种病原体，以便卫生工作者能够开始最适当的治疗，而不会产生不必要的耐药病原体风险。为了鼓励全球范围内的采用和使用，该测试也将易于使用，非常稳定，具有长期的现场部署效力，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且功率密集的制冷以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还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，即使是运行免疫测定的最基本要求，即电力、清洁水和培训，在教育、资源和社会基础设施最少的发展中地区也是极其昂贵或不可能的。在第一阶段，内部监督办公室将开发一种侧流免疫测定（LFIA）试纸，能够在一个样本中检测多种传染性病原体，假阳性或假阴性发生率低。UST的专利技术将保护试纸，即使在低于50 ℃的温度下长时间储存两年，也能保持完整的活性。我们将建立一个简单的，一步到位的方案，使最低限度的培训工作人员可以产生准确和一致的结果，即使是原始粪便样本。使用高荧光量子点（QD）探针，可以用肉眼检测，这种敏感的系统将在15分钟内产生样品中存在的病原体的定性结果。在第二阶段，IOS将把更多的其他粪便肠道病原体的免疫检测方法整合到同一种试纸上，以进行全面的病原体筛查。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，便携式和坚固的试纸阅读器，用于增强定量检测。在第二阶段，监督厅还将准备材料，列入FDA批准申请，以便对原型系统进行临床评估。公共卫生相关性：每年，腹泻导致数百万人死亡，特别是五岁以下的儿童。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。在世界上许多经济上、气候上和/或地理上具有挑战性的地区，由于其蛋白质试剂的脆弱性质和进行测定的技术困难，灵敏的免疫测定诊断是不可用的。一种使用简单、超稳定的试纸系统，可以准确区分单个样本中的几种潜在病原体，这将有助于适当的医疗干预，并提供确定的治疗方案。多路复用格式还将消除对多次测试的需要，降低单位成本，并扩大对这些关键工具的访问，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.