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:
    8044040
  • 负责人:
  • 金额:
    $ 20万
  • 依托单位:
  • 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

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):该项目的目标是将等温放大系统与横向流动检测相结合,以生产一种简单、灵敏、低成本的一体式黄病毒检测设备,该设备易于在护理环境中执行和解释,并适合在服务较差的社区使用。黄病毒科的病毒,如登革热、西尼罗河病毒和日本脑炎,是昆虫传播的病毒,对人类和兽医都很重要。根据世卫组织的数据,仅登革热病毒一项就影响了全球120万人,导致1998年3442人死亡。这些疾病通过蚊子和扁虱传播,在自然界中通常处于传播周期。它们广泛分布在世界各地,尽管一种特定的黄病毒可能在地理上受到限制。它们在人类身上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,但许多病毒尚未被证明会导致人类疾病。它们在相似的地理区域出现重叠的临床症状和体征。目前,这些感染的诊断速度慢、费用高、劳动密集型,延误诊断往往会导致严重的后遗症,有时甚至死亡。因此,我们建议开发一种针对几种黄病毒的全面检测方法,最初的重点是登革热病毒和西尼罗河病毒。这些测试是基于等温核酸放大技术。我们将比较已有的环路介导扩增(LAMP)技术与我们新开发的SPIDR方法,后者在早期研究中似乎更容易实现,并提供更高的特异性。SPIDR和LAMP都是等温技术,不需要热循环仪,只需不到30分钟就能得出结果。扩增产物将在使用横向流动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。一旦开发出一种测试,它将非常容易执行,并且不需要在样品制备时进行DNA分离(与PCR相比)。这种简化的检测便于技术较差的操作员进行解释。开发后,该设备将不需要仪器或训练有素的科学家来操作,特别适合资源匮乏的环境。该项目是由PI与几位有成就的科学家合作提出的,涉及广泛的协同研究兴趣领域。这项拟议的研究将解决一个基本的、与项目相关的需求,即对发展中国家以及农村和社会经济贫困的美国人有用的关注点测试。 与公共卫生相关:该项目的目标是完全集成一种简单、低成本、一体化的黄病毒检测扩增和检测设备,该设备可以在资源匮乏的环境中以关怀的方式轻松执行和解释。该设备将不需要仪器或 训练有素的科学家操作。开发一种可靠、灵敏和特异的血液样本和其他体液中黄病毒的“关注点”检测方法,将是许多低资源环境的重大突破。

项目成果

期刊论文数量(0)
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DAVID Alan MEAD其他文献

DAVID Alan MEAD的其他文献

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{{ truncateString('DAVID Alan MEAD', 18)}}的其他基金

High Fidelity Linear MicroVector to Clone Complex, Problematic, and Large DNAs
用于克隆复杂、有问题和大型 DNA 的高保真线性微载体
  • 批准号:
    9346284
  • 财政年份:
    2017
  • 资助金额:
    $ 20万
  • 项目类别:
Expression Enhanced Natural Product Pathways Using Advanced Metagenomic Tools
使用先进的宏基因组工具表达增强的天然产物途径
  • 批准号:
    8903016
  • 财政年份:
    2015
  • 资助金额:
    $ 20万
  • 项目类别:
Analytical Metagenomics Paradigm for Structure Based Screening
基于结构的筛选的分析宏基因组学范式
  • 批准号:
    8310684
  • 财政年份:
    2012
  • 资助金额:
    $ 20万
  • 项目类别:
Enabling Technologies for Low Resource Molecular Diagnostics
低资源分子诊断的支持技术
  • 批准号:
    7779226
  • 财政年份:
    2010
  • 资助金额:
    $ 20万
  • 项目类别:
NOVEL SOLUBILITY-ENHANCING PROTEIN EXPRESSION TECHNOLOGY
新型增强溶解度的蛋白质表达技术
  • 批准号:
    8501542
  • 财政年份:
    2009
  • 资助金额:
    $ 20万
  • 项目类别:
NOVEL SOLUBILITY-ENHANCING PROTEIN EXPRESSION TECHNOLOGY
新型增强溶解度的蛋白质表达技术
  • 批准号:
    8250969
  • 财政年份:
    2009
  • 资助金额:
    $ 20万
  • 项目类别:
Massively Parallel Single Cell Genomics of the Human Microbiome
人类微生物组的大规模并行单细胞基因组学
  • 批准号:
    7611525
  • 财政年份:
    2008
  • 资助金额:
    $ 20万
  • 项目类别:
HIGH THROUGHPUT CLONING OVEREXPRESSION AND PURIFICATION OF ACTIVE MEMBRANE PROT
活性膜保护的高通量克隆过表达和纯化
  • 批准号:
    7325336
  • 财政年份:
    2007
  • 资助金额:
    $ 20万
  • 项目类别:
Ex Cyto DNA Sequencing from Single Bacterial Colonies
单菌落的前细胞 DNA 测序
  • 批准号:
    7870519
  • 财政年份:
    2006
  • 资助金额:
    $ 20万
  • 项目类别:
Ex Cyto DNA Sequencing from Single Bacterial Colonies
单菌落的前细胞 DNA 测序
  • 批准号:
    7156326
  • 财政年份:
    2006
  • 资助金额:
    $ 20万
  • 项目类别:

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