Strain Specific Detection of Influenza at the Point-of-Care

在护理点对流感病毒株进行特异性检测

基本信息

项目摘要

DESCRIPTION (provided by applicant): Strain Specific Detection of Influenza at the Point-of-Care 1. PROJECT SUMMARY Influenza viruses cause significant morbidity and mortality worldwide; World Health Organization (WHO) estimates 250,000 ~ 500,000 yearly deaths. In particular, a highly pathogenic strain - avian influenza virus H5N1 (15, 56) - poses a threat of a possible pandemic because it can rapidly mutate to acquire the ability to transmit among humans (23, 49). The detection of minimal human infectious dose (~100 viral particles) (65) of Influenza A from patient samples (e.g. nasal swabs or mucus) is a significant analytical challenge. Detection by viral culture is sensitive and specific; however, this method is slow (3-10 days) and cannot be performed at POC. Surface marker-based detection (e.g. ELISA) provide a simple and rapid testing, however, it suffers from a poor limit of detection (~ 105 viral particles/ml) (9, 21, 59). Currently, polymerase chain reaction (PCR) offers the highest sensitivity in a much shorter turn-around time (2-4 hours) (29, 61, 67). Nevertheless, due to the difficulties in POC operation, patient samples are currently analyzed at central laboratories which typically requires ~2-3 days. Currently in the field, there is no automated technology solution that can perform specific influenza detection directly from patient samples with high sensitivity and specificity, and the WHO has specifically emphasized the critical need to fill this important technological gap (75). Towards a solution to this important problem, this group of three PI's with complementary skill sets (influenza virology, aptamer biochemistry and microfluidic device engineering) propose to develop a powerful, field- portable, microfluidic platform for Influenza detection at POC. This effort is truly unique in many facets; first, to circumvent the problem posed by rapid mutation of the coat protein, we propose to generate novel DNA aptamer reagents which will specifically bind to the nucleoprotein complex of the virus. Currently, there are no reported aptamers for the nucleoprotein complex, and due to the fact that the nucleoprotein is conserved, it will serve as a universal tag to label Influenza A. Secondly, the aptamer reagent will be conjugated to magnetic beads to purify the nucleoprotein complex from nasal swab samples using our unique micromagnetics technology. Thirdly, by leveraging our expertise in miniaturized genetic analysis systems, we will develop the IMED chip capable of 1) integrated micromagnetic sample purification, 2) reverse transcription-PCR amplification and 3) sequence-specific electrochemical detection in a single monolithic device. The successful completion of this project will allow a quantitative "sample-to-answer" capability - the input into the system will be unprocessed patient samples, and the output will be an electrochemical signal that will be directly proportional to the number of copies of specific influenza RNA sequences in the sample. Such powerful combination of novel affinity reagents with highly integrated disposable devices will enable the development of critically needed effective POC diagnostics systems. PUBLIC HEALTH RELEVANCE: Strain Specific Detection of Influenza at the Point-of-Care Due to the fact that the envelope of influenza A virus differs among subtypes and evolves continuously, there is an urgent need for a field-portable genetic detection platform that can rapidly identify pathogenic strain of viral pathogens from unprocessed samples with high sensitivity and specificity. Towards an effective solution, the three PI's with complementary skill sets (influenza virology, aptamer biochemistry and microfluidic device engineering) propose to develop an effective point-of-care diagnostic system for influenza by 1) generating specific, high affinity DNA aptamers to tag the conserved regions of Influenza A, and 2) developing a highly integrated microfluidic system capable of magnetic sample preparation, RT-PCR amplification and sequence specific electrochemical detection in a single chip. By integrating the functions in a monolithic chip, the success of this project will yield a novel POC analysis system with unprecedented capabilities which will have a significant impact for Influenza A detection as well as many other applications in food safety, environmental monitoring and homeland security. 1
描述(由申请方提供):床旁流感病毒菌株特异性检测1。流感病毒在世界范围内引起显著的发病率和死亡率;世界卫生组织(WHO)估计每年有250,000 ~ 500,000人死亡。特别是,高致病性毒株--禽流感病毒H5 N1(15,56)--构成了可能发生大流行的威胁,因为它可以迅速变异以获得在人类之间传播的能力(23,49)。从患者样本(例如鼻拭子或粘液)中检测最小人类感染剂量(约100个病毒颗粒)(65)的甲型流感病毒是一项重大的分析挑战。通过病毒培养进行检测具有灵敏度和特异性;然而,该方法速度较慢(3-10天),无法在POC下进行。基于表面标志物的检测(例如ELISA)提供了简单和快速的检测,然而,其存在检测限差(~ 105病毒颗粒/ml)的问题(9,21,59)。目前,聚合酶链反应(PCR)在更短的周转时间(2-4小时)内提供了最高的灵敏度(29,61,67)。然而,由于POC操作的困难,目前在中心实验室分析患者样本,通常需要约2-3天。目前在该领域,没有自动化技术解决方案可以直接从患者样本中进行高灵敏度和特异性的特异性流感检测,WHO特别强调了填补这一重要技术空白的迫切需要。为了解决这一重要问题,具有互补技能集(流感病毒学、适体生物化学和微流体装置工程)的这三个PI的组提出开发用于POC的流感检测的强大的、现场便携的微流体平台。这一努力在许多方面都是真正独特的;首先,为了避免外壳蛋白快速突变所带来的问题,我们建议产生新的DNA适体试剂,它将特异性地结合到病毒的核蛋白复合物上。目前,还没有报道用于核蛋白复合物的适体,并且由于核蛋白是保守的,它将作为标记甲型流感的通用标签。其次,将适体试剂与磁珠结合,使用我们独特的微磁性技术从鼻拭子样本中纯化核蛋白复合物。第三,通过利用我们在小型化遗传分析系统方面的专业知识,我们将开发能够在单个单片设备中实现1)集成微磁样品纯化,2)逆转录PCR扩增和3)序列特异性电化学检测的IMED芯片。该项目的成功完成将允许定量的“样本到答案”能力-输入系统的将是未处理的患者样本,输出将是电化学信号,该信号将与样本中特定流感RNA序列的拷贝数成正比。新型亲和试剂与高度集成的一次性装置的这种强大组合将能够开发急需的有效POC诊断系统。公共卫生相关性:由于甲型流感病毒的包膜在不同亚型之间存在差异并不断进化,因此迫切需要一种现场便携式基因检测平台,该平台可以从未经处理的样本中快速识别病毒病原体的致病株,具有高灵敏度和特异性。为了有效的解决方案,三个具有互补技能的PI(流感病毒学、适体生物化学和微流体装置工程)提出通过以下方式开发用于流感的有效的即时诊断系统:1)产生特异性、高亲和力DNA适体以标记甲型流感的保守区域,和2)开发能够进行磁性样品制备的高度集成的微流体系统,RT-PCR扩增和序列特异性电化学检测在一个芯片。通过将这些功能集成到单片芯片中,该项目的成功将产生一种具有前所未有的功能的新型POC分析系统,这将对甲型流感检测以及食品安全,环境监测和国土安全等许多其他应用产生重大影响。1

项目成果

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HYONGSOK Tom SOH其他文献

HYONGSOK Tom SOH的其他文献

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{{ truncateString('HYONGSOK Tom SOH', 18)}}的其他基金

Real-time biosensor for mapping the function of the pancreas
用于绘制胰腺功能的实时生物传感器
  • 批准号:
    9926632
  • 财政年份:
    2019
  • 资助金额:
    $ 39.47万
  • 项目类别:
Real-time biosensor for mapping the function of the pancreas
用于绘制胰腺功能的实时生物传感器
  • 批准号:
    10455342
  • 财政年份:
    2018
  • 资助金额:
    $ 39.47万
  • 项目类别:
Real-time biosensor for mapping the function of the pancreas
用于绘制胰腺功能的实时生物传感器
  • 批准号:
    10170068
  • 财政年份:
    2018
  • 资助金额:
    $ 39.47万
  • 项目类别:
Micromagnetic Aptamer PCR System for Ultrasensitive Multiplexed Protein Detection
用于超灵敏多重蛋白质检测的微磁适体 PCR 系统
  • 批准号:
    7812154
  • 财政年份:
    2009
  • 资助金额:
    $ 39.47万
  • 项目类别:
Strain Specific Detection of Influenza at the Point-of-Care
在护理点对流感病毒株进行特异性检测
  • 批准号:
    8272670
  • 财政年份:
    2009
  • 资助金额:
    $ 39.47万
  • 项目类别:
Strain Specific Detection of Influenza at the Point-of-Care
在护理点对流感病毒株进行特异性检测
  • 批准号:
    8070475
  • 财政年份:
    2009
  • 资助金额:
    $ 39.47万
  • 项目类别:
Strain Specific Detection of Influenza at the Point-of-Care
在护理点对流感病毒株进行特异性检测
  • 批准号:
    7917202
  • 财政年份:
    2009
  • 资助金额:
    $ 39.47万
  • 项目类别:
Micromagnetic Aptamer PCR System for Ultrasensitive Multiplexed Protein Detection
用于超灵敏多重蛋白质检测的微磁适体 PCR 系统
  • 批准号:
    7641885
  • 财政年份:
    2009
  • 资助金额:
    $ 39.47万
  • 项目类别:
Rapid Generation of High Affinity Protein Sensor Elements
快速生成高亲和力蛋白质传感器元件
  • 批准号:
    7569990
  • 财政年份:
    2008
  • 资助金额:
    $ 39.47万
  • 项目类别:
Rapid Generation of High Affinity Protein Sensor Elements
快速生成高亲和力蛋白质传感器元件
  • 批准号:
    7472611
  • 财政年份:
    2008
  • 资助金额:
    $ 39.47万
  • 项目类别:

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基于石墨烯适体技术的压力监测即时诊断
  • 批准号:
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