Enabling Technologies for Low Resource Molecular Diagnostics

低资源分子诊断的支持技术

• 批准号: 8044040
• 负责人: DAVID Alan MEAD
• 金额: $ 20万
• 依托单位: LUCIGEN CORPORATION
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044040
• 关键词: Address; Affect; American; Arboviruses; Area; Bedside Testings; Biological Assay; Blood specimen; Body Fluids; Caring; Cessation of life; Clinical; Collaborations; Communities; Complex; Culicidae; DNA; Dengue; Dengue Virus; Detection; Developing Countries; Development; Devices; Diagnosis; Diagnostic; Disadvantaged; Disease; Economics; Encephalitis; Family; Flaviviridae; Flavivirus; Gender; Genetic Variation; Geographic Locations; Goals; Health; Health Services Accessibility; Health behavior; Hour; Human; Individual; Infection; Infectious Agent; Insecta; Japanese Encephalitis; Japanese Encephalitis Viruses; Japanese encephalitis virus; Lateral; Life; Mediating; Methods; Minority Groups; Molecular; Nature; Nucleic Acids; Phase; Preparation; Process; Quality of Care; Race; Research; Resources; Rural; Sampling; Scientist; Signs and Symptoms; Small Business Innovation Research Grant; Specificity; Specimen; System; Techniques; Technology; Testing; Ticks; Training; United States; Veterinary Medicine; Viral Hemorrhagic Fevers; Virus; Visual; West Nile virus; base; cost; global health; instrumentation; interest; point of care; programs; public health relevance; response; social health determinants; socioeconomics; transmission process

DESCRIPTION (provided by applicant): The goal of this project is to integrate an isothermal amplification system with lateral flow detection to produce a simple, sensitive, low cost, all-in-one detection device for flaviviruse that is easily performed and interpreted in a point of care setting and is amenable to use in poorly served communities. The viruses of the family Flaviviridae, e.g. dengue, West Nile, and Japanese Encephalitis, are insect-borne viruses important to both human and veterinary medicine. According to WHO, dengue virus alone affected 1.2 million globally, resulting in 3442 deaths in 1998. These diseases are transmitted by mosquito and ticks and usually are maintained in a transmission cycle in nature. They are widely distributed throughout the world, although a specific flavivirus may be geographically restricted. They produce a broad spectrum of clinical responses in humans ranging from asymptomatic infection to fulminant encephalitis or haemorrhagic fever. Nearly 60 flaviviruses are known to exist, although but many are yet to be shown to cause disease in humans. They present with overlapping clinical signs and symptoms in similar geographic areas. The diagnosis of these infections is currently slow, expensive, and labor intensive and a delay in diagnoses often leads to serious sequelae and sometimes death. Thus, we propose to develop a comprehensive assay specific for several flaviviruses, with initial focus on dengue virus and West Nile virus. These tests are based on isothermal nucleic acid amplification technologies. We will compare the established technology, Loop-mediated AMPlification (LAMP), to our newly developed method SPIDR, which in early studies appears to be easier to implement and affords greater specificity. Both SPIDR and LAMP are isothermal techniques, precluding the need for thermocyclers and can produce results in less than 30 minutes. The amplification products will be processed in an integrated system using lateral flow technology, allowing sensitive, rapid, unambiguous detection with the potential for multiplexing. Once a test is developed, it will be very simple to perform and does not require DNA isolation for sample preparation (as compared to PCR). Such simplified detection facilitates interpretation by less skilled operators. When developed, the device will not require instrumentation or trained scientists to operate and is especially suitable to resource poor settings. The project is proposed by the PI in collaboration with several accomplished scientists across a broad field of synergistic research interests. The proposed research will address a fundamental, program-relevant need for a point of care test useful in developing country settings as well as for rural and socioeconomically disadvantaged Americans. PUBLIC HEALTH RELEVANCE: The goal of this project is to fully integrate a simple, low cost, all-in-one amplification and detection device for flavivirus testing that is easily performed and interpreted in a point of care manner in resource poor settings. The device will not require instrumentation or trained scientists to operate. The development of a reliable, sensitive and specific "point of care" test for flaviviruses in blood samples and other body fluids would be a major breakthrough for many low resource settings.

描述（由申请人提供）：本项目的目标是将等温扩增系统与侧流检测相结合，以产生一种简单、灵敏、低成本、一体化的黄嘌呤检测装置，该装置易于在护理点环境中进行和解释，并且适合在服务较差的社区使用。黄病毒科的病毒，例如登革热、西尼罗河和日本脑炎，是对人类和兽医学都很重要的虫媒病毒。根据世界卫生组织的统计，1998年，仅登革热病毒就影响了全球120万人，导致3442人死亡。这些疾病由蚊子和蜱传播，通常在自然界中保持传播周期。它们广泛分布于世界各地，尽管特定的黄病毒可能受到地理限制。它们在人类中产生广泛的临床反应，从无症状感染到暴发性脑炎或出血热。已知存在近60种黄病毒，尽管其中许多尚未被证明会引起人类疾病。他们在相似的地理区域表现出重叠的临床体征和症状。这些感染的诊断目前是缓慢的，昂贵的，劳动密集型和诊断的延误往往会导致严重的后遗症，有时甚至死亡。因此，我们建议开发一种全面的检测特异性的几种黄病毒，最初的重点是登革热病毒和西尼罗河病毒。这些测试基于等温核酸扩增技术。我们将比较已建立的技术，环介导的AMP化（LAMP），我们新开发的方法SPIDR，在早期研究中似乎更容易实施，并提供更大的特异性。SPIDR和LAMP都是等温技术，不需要热循环仪，并且可以在不到30分钟内产生结果。扩增产物将在使用侧流技术的集成系统中进行处理，从而实现灵敏、快速、明确的检测，并具有多路复用的潜力。一旦开发了测试，它将非常容易执行，并且不需要DNA分离用于样品制备（与PCR相比）。这种简化的检测便于不太熟练的操作者进行解释。当开发出来时，该设备将不需要仪器或训练有素的科学家来操作，特别适合资源贫乏的环境。该项目是由PI与几位有成就的科学家在广泛的协同研究领域合作提出的。拟议的研究将解决一个基本的，与计划相关的需要，在发展中国家的设置以及农村和社会经济上处于不利地位的美国人有用的护理点测试。 公共卫生关系：该项目的目标是完全集成一个简单，低成本，多合一的扩增和检测设备，用于黄病毒检测，易于在资源匮乏的环境中以护理点的方式进行和解释。该器械不需要手术工具， 训练科学家来操作。开发一种可靠、敏感和特异的“即时”检测血液样本和其他体液中的黄病毒的方法，将是许多资源匮乏环境的一项重大突破。