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

用于肠道病原体多重检测的超稳定试纸系统

• 批准号: 7404692
• 负责人: VLADIMIR RUBTSOV
• 金额: $ 15.57万
• 依托单位: INTELLIGENT OPTICAL SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-15 至 2009-11-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7404692
• 关键词: Address; Adoption; Age; Alloys; Antibiotic Resistance; Antibodies; Bacteria; Bedside Testings; Biological Assay; Biological Preservation; Cause of Death; Cessation of life; Child; Collaborations; Communicable Diseases; Condition; Cryptosporidium; Cryptosporidium parvum; Detection; Diagnosis; Diagnostic; Diagnostic tests; Diarrhea; Education; Environment; Epidemic; Escherichia coli EHEC; Escherichia coli O157; Evaluation; Exhibits; Expert Systems; Eye; Feces; Goals; Government; Hand; Health; Hour; Immunoassay; Immunological Diagnosis; Incidence; Individual; Infant; Intervention; Lateral; Left; Legal patent; Medical; Membrane; Nature; One-Step dentin bonding system; Oocysts; Optics; Parasites; Persons; Phase; Process; Propionibacterium acnes; Proteins; Protocols documentation; Public Health; Purpose; Quantum Dots; Reader; Reagent; Refrigeration; Research Infrastructure; Resources; Risk; Running; Sampling; Sensitivity and Specificity; Survival Rate; System; Technology; Temperature; Testing; Therapeutic Corynebacterium Parvum; Training; United States Food and Drug Administration; Virus; Water; Work; cost; cross reactivity; day; design; enteric pathogen; experience; fungus; improved; innovation; microorganism; pathogen; point of care; prevent; prototype; social; tool; user-friendly

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.

描述（由申请人提供）：腹泻是全球主要死亡原因，尤其是医疗资源有限的婴幼儿。尽管准确及时地诊断致病传染病将提高存活率并减轻流行病传播，但目前在不可控环境中运输和储存敏感诊断材料的挑战在经济或后勤上并不可行。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 (UST) 提议创建一种超稳定的一步式免疫分析平台，能够以高灵敏度和特异性识别单个样品中的多种肠道病原体。这种快速且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启动最合适的治疗，而不会产生产生抗生素抗性病原体的不必要风险。为了鼓励全球范围内的采用和使用，该测试还将易于使用、非常稳定、现场部署效果长、敏感且针对微生物且价格低廉。现有的免疫诊断在该领域是不足且不切实际的，因为它们通常需要昂贵且耗电的制冷才能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，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最少的发展中地区，即使是进行免疫测定的最基本要求，即电力、清洁水和培训，也极其昂贵或不可能。在第一阶段，IOS将开发一种侧流免疫分析（LFIA）测试条格式，能够检测单个样本中的多种传染性病原体，且假阳性或阴性发生​​率低。 UST 的专利技术将保存测试条，以便即使在 <50oC 的温度下长期储存两年后仍能保留全部活性。我们将建立一种简单的一步式方案，以便经过最低限度培训的工作人员即使使用原始粪便样本也能产生准确且一致的结果。该敏感系统使用肉眼可检测的高荧光量子点 (QD) 探针，可在 15 分钟内得出样品中病原体的定性结果。在第二阶段，IOS 将把更多针对其他粪便肠道病原体的免疫测定集成到相同的测试条格式上，以进行全面的病原体筛查。对于在低病原体浓度下具有极强传染性的传染病，我们将构建一种小型、廉价、便携式且坚固的测试条阅读器，以增强定量检测。在第二阶段，IOS还将准备材料纳入FDA批准申请中，以便原型系统可以进行临床评估。公共卫生相关性：每年，腹泻都会导致数百万人死亡，尤其是五岁以下的儿童。许多病原体，包括细菌、寄生虫、病毒和真菌，可能在短短 72 小时内引起致命性腹泻。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以及进行测定的技术难度，敏感的免疫测定诊断是不可用的。一种易于使用且超稳定的测试条系统，可以准确地区分单个样本中的几种潜在病原体，将有助于适当的医疗干预并提供确定的治疗选择。多路复用格式还可以消除多次测试的需要，降低单位成本，并扩大对这些关键工具的访问范围，以防止不必要的死亡。