Enabling Technologies for Low Resource Molecular Diagnostics

低资源分子诊断的支持技术

基本信息

批准号：
7779226
负责人：
DAVID Alan MEAD
金额：
$ 20万
依托单位：
LUCIGEN CORPORATION
依托单位国家：
美国
项目类别：
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/7779226
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

项目摘要

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）：该项目的目的是将等温放大系统集成具有横向流量检测的等温放大系统，以生成一种简单，敏感，低成本的，用于Flaviviruse的全合一检测设备，在护理设置中易于执行和解释，并且可以在服务不善的社区中使用。 Flaviviridae家族的病毒，例如登革热，西尼罗河和日本脑炎是对人类和兽医医学都很重要的昆虫传播病毒。根据WHO的说法，仅登革热病毒在全球范围内影响了120万，1998年导致3442例死亡。这些疾病是由蚊子和壁虱传播的，通常在自然界中保持传播周期。尽管特定的黄病毒可能受到地理位置的限制，但它们在世界范围内广泛分布。它们在人类中产生各种临床反应，从无症状感染到暴发性脑炎或出血热。已知存在近60个黄病毒，但尚未证明许多人在人类中引起疾病。它们在类似地理区域中出现重叠的临床体征和症状。这些感染的诊断目前缓慢，昂贵且劳动密集型和诊断延迟通常会导致后遗症严重，有时会导致死亡。因此，我们建议开发针对几种黄病毒的全面测定，最初关注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。我们将将既定的技术，循环介导的放大（LAMP）与我们新开发的方法SPIDR进行比较，SPIDR在早期研究中似乎更容易实施并提供更高的特异性。 SPIDR和LAMP都是等温技术，排除了对热循环器的需求，并且可以在不到30分钟的时间内产生结果。扩增产物将使用横向流技术在集成系统中处理，从而允许敏感，快速，明确的检测，并具有多元化的潜力。一旦开发了测试，它将非常易于执行，并且不需要DNA进行样品制备（与PCR相比）。这种简化的检测有助于熟练的操作员的解释。开发后，该设备将不需要仪器或训练有素的科学家进行操作，并且特别适合资源不良的设置。 PI与在广泛的协同研究兴趣领域的几位有成就的科学家合作提出了该项目。拟议的研究将解决针对发展中国家环境以及农村和社会经济处境不利的美国人的基本且与计划的基本需求。公共卫生相关性：该项目的目的是完全整合一项简单，低成本的放大和检测设备，以用于Flavivirus测试，该设备在资源较差的设置中易于执行和解释。该设备将不需要仪器或训练有素的科学家进行操作。对于许多低资源设置，对血液样本和其他体液中黄病毒的可靠，敏感和特定的“护理点”测试的开发将是一个重大突破。