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; 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）：腹泻是全球死亡的主要原因，尤其是在获得医疗资源有限的婴儿和幼儿中。尽管对病因传染病的准确及时诊断会增加存活率并减轻流行病的差异，但在无法控制的环境中运输和存储敏感诊断的当前挑战在不可控制的环境中并不经济或逻辑上可行。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）建议创建一个超稳的一步免疫测定平台，该平台可以在具有高灵敏度和特异性的单个样本中识别多个肠道病原体。这种快速且廉价的护理测试将准确识别一种或多种病原体，以便卫生工作者可以发起最合适的治疗方法，而不会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，该测试也将易于使用，非常稳定，具有长期的现场部署功效，特定于敏感和微生物，并且便宜。现有的免疫诊断在该领域不足和不切实际，因为它们通常需要昂贵且功能密集型的冷藏以在无法控制的环境条件下保留疗效。此外，许多免疫测定的有效执行还需要使用多步协议的培训和经验，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，在开发最少的教育，资源和社会基础设施的发展地区中，也非常昂贵或不可能。在第一阶段，iOS将开发出横向流量免疫测定（LFIA）测试条格式，能够在单个样本中检测出多种传染病病原体，该样品的发生率较低。 UST的专利技术将保留测试条，以便在两年内延长<50oC的温度延长储存后，可以保留全部活动。我们将建立一个简单的一步协议，以便训练有素的工人即使使用原始的粪便样本也可以产生准确，一致的结果。使用可在肉眼中检测到的高荧光量子点（QD）探针，该敏感系统将在15分钟内产生样品中存在的病原体的定性结果。在II阶段，iOS将将更多的其他粪肠病毒的免疫分析集成到同一测试带格式中，以产生全面的病原体筛查。对于在低病原体浓度下具有极大传染性的传染病，我们将建立一个小型，廉价，便携式和坚固的测试带读取器，以增强定量检测。在第二阶段，iOS还将准备材料，以包括在FDA批准的应用中，以便可以对原型系统进行临床评估。公共卫生相关性：每年，腹泻会导致数百万人，尤其是五岁以下的儿童死亡。包括细菌，寄生虫，病毒和真菌在内的许多病原体可能在72小时内引起致命的腹泻。在许多经济上，从气候学和/或世界上具有挑战性的地区，敏感的免疫测定诊断无法获得，因为其蛋白质试剂的脆弱性质以及进行测定的技术困难。一种易于使用和超稳定的测试带系统，可以准确区分单个样品中的几种潜在病原体，将有助于适当的医疗干预措施，并确定治疗选择。多路复用格式还将消除对多次测试的需求，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。