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

描述(申请人提供)：腹泻是全球主要的死亡原因，特别是在获得医疗资源有限的婴幼儿中。尽管对致病传染病的准确和及时诊断将提高存活率并减缓疫情传播，但目前在无法控制的环境中运输和储存敏感诊断信息的挑战在经济上或后勤上都是不可行的。为了满足这一需求，智能光学系统(IOS)和合作者通用稳定技术公司(UST)提出创建一个超稳定的一步免疫分析平台，可以在一个样本中识别多个肠道病原体，具有高灵敏度和高特异性。这一快速且廉价的护理点检测将准确地识别一种或多种病原体，以便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性病原体的不必要风险。为了鼓励全球采用和使用，这种测试也将易于使用，非常稳定，现场部署长期有效，敏感性和微生物特异性，并且价格低廉。现有的免疫诊断技术在现场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效力。此外，许多免疫分析的有效执行还需要培训和使用多步骤方案的经验，才能产生有意义的结果。不幸的是，在教育、资源和社会基础设施极少的发展中地区，即使是运行免疫分析的最基本要求，即电力、清洁水和培训，也昂贵得令人望而却步，甚至是不可能的。在第一阶段，iOS将开发一种侧向流动免疫分析(LFIA)试纸格式，能够在一个样本中检测多种感染性病原体，假阳性或假阴性的发生率较低。科大的专利技术将保留试条，这样即使在50摄氏度的温度下延长两年的储存时间，也能保持全部活性。我们将建立一种简单的一步法，这样经过最低限度培训的工作人员可以产生准确和一致的结果，即使是原始的粪便样本。使用肉眼可检测到的高荧光量子点(QD)探针，这个灵敏的系统将在15分钟内对样品中存在的病原体做出定性结果。在第二阶段，iOS将把更多对其他粪便肠道病原体的免疫分析整合到相同的试纸格式中，以产生全面的病原体筛查。对于在低病原体浓度下极具传染性的传染病，我们将建造一种小型、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小时内导致致命的腹泻。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易碎性和进行分析的技术难度，无法获得敏感的免疫分析诊断方法。一种简单易用、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，并提供一定的治疗选择。多元化的格式还将消除多次测试的需要，降低单位成本，并扩大这些关键工具的使用范围，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.