Species Level Identification of Pathogenic Viruses on a Microfluidic Platform

微流控平台上病原病毒的物种水平鉴定

• 批准号: 7291673
• 负责人: LOUIS H STRONG
• 金额: $ 40.49万
• 依托单位: RADIATION MONITORING DEVICES, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-15 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7291673
• 关键词: Adopted; Aerosols; Antibodies; Architecture; Area; Base Sequence; Binding; Biological; Biological Assay; Biosensor; Birefringence; Capsid Proteins; Clinical; Collection; Condition; Cytolysis; Data Analyses; Detection; Development; Diffusion; Drops; Emergency Situation; Engineering; Excision; Exhibits; Fluorescence; Funding; Gene Targeting; Genes; Genetic Markers; Genetic Transcription; Genome; Genomics; Health Sciences; Human Resources; Label; Libraries; Light; Lighting; Link; Measurable; Measurement; Measures; Medical Surveillance; Methodology; Methods; Microfluidics; Modeling; Molecular; Monitor; Nanosphere; Nucleic Acids; Nucleotides; Numbers; Optics; Pathogenicity; Performance; Phase; Photons; Preparation; Probability; Problem Solving; Process; Property; Proteins; Protocols documentation; Quarantine; RNA; RNA Viruses; RNA amplification; Radiation Monitoring; Rate; Reaction; Reagent; Recombinants; Relaxation; Reporter; Sampling; Schedule; Scheme; Self-Sustained Sequence Replication; Sensitivity and Specificity; Series; Signal Transduction; Spectrum Analysis; Speed; Staging; Structure; System; Temperature; Tennessee; Testing; Time; Transfection; Universities; Vesicular stomatitis Indiana virus; Viral; Viral Genes; Viral Genome; Viral Load result; Virion; Virulent; Virus; air monitoring; base; design; detector; innovative technologies; instrument; instrumentation; microchip; monitoring device; mutant; nanoparticle; novel; parallel processing; particle; pathogen; performance tests; prototype; sample collection; sensor; viral detection; virus envelope; wasting; waterborne

DESCRIPTION (provided by applicant): Radiation Monitoring Devices, Inc. (RMD) proposes to develop a field-portable instrument for the detection and identification of pathogenic viruses that are collected by an aerosol concentrator. It will incorporate modular, microfluidic platforms designed to capture and concentrate the sample for a rapid, two-stage analysis. The first employs pathogen capture on antibody-decorated, paramagnetic nanospheres and will provide rapid, preliminary warning based on the hydrodynamic properties of the nanoparticles when the targeted viruses are bound. This stage will monitor the relaxation of the magneto-optical birefringence of the nanoparticles and will trigger an alarm when the birefringence relaxation rate drops off dramatically. The methodology allows for continuous surveillance over virtually unlimited periods. The second stage will provide either confirmation or correction to the preliminary identification. It is based on the ability to interrogate the captured nucleic acid with a series of unique probes that change their fluorescence properties when they hybridize with specific nucleotide sequences of suspected viral genes. We will compare the speed and sensitivity of detecting these genes by fluorescence generated from molecular beacons, and by the temporal correlation of fluorescence issued from tagged probes. The detection method exhibiting the best performance will be incorporated into the sensor. In each case, Nucleic Acid Sequence Based Amplification (NASBA), an isothermal nucleotide amplification scheme, will be employed to raise the number of target template copies to detectable levels. The biosensor will integrate the two stages of sample transfer, amplification, and analysis in a microfluidic architecture that enables high throughput processing and parallel detection of multiple probes. Employing both proportional mode avalanche photodiode (APD) detectors and Geiger-mode muAPDs that are responsive to extremely low light levels, sensitivity to low virus titers will be possible. This proposal solicits funding for developing an instrument to detect and identify pathogenic viruses in a rapid, automated two-step analysis that can be used by emergency workers who are responding to a suspected release of airborne (waterborne) agents. The first step screens the sample to determine if there are any virus particles that possess a targeted group of exposed coat proteins. If the answer is affirmative, the virus is isolated and its nucleic acid is analyzed to determine if it carries pathogen genes.

描述（由申请人提供）：辐射监测设备，Inc。（RMD）提议开发一种可检测和鉴定由气溶胶浓度收集的病毒病毒的可供货运仪器。它将结合模块化的微流体平台，旨在捕获和集中样品进行快速的两阶段分析。第一个使用病原体捕获在抗体装饰的，顺磁性纳米球上，并将在靶向病毒结合时根据纳米颗粒的流体动力特性提供快速的初步警告。此阶段将监视纳米颗粒的磁光双折射的松弛，并在双折射放松率急剧下降时触发警报。该方法允许在几乎无限的时期内进行连续监视。第二阶段将为初步识别提供确认或校正。它基于使用一系列独特的探针询问捕获的核酸的能力，这些探针与疑似病毒基因的特定核苷酸序列杂交时会改变其荧光性能。我们将通过从分子信标产生的荧光以及通过标记探针发出的荧光的时间相关性来比较检测这些基因的速度和灵敏度。表现出最佳性能的检测方法将纳入传感器。在每种情况下，核酸序列的放大（NASBA）（一种等温核苷酸扩增方案）将采用将目标模板副本的数量提高到可检测水平。生物传感器将在微流体体系结构中整合样品转移，扩增和分析的两个阶段，从而实现高吞吐量处理和对多个探针的平行检测。采用比例模式雪崩光电二极管（APD）检测器和吉格模式MUAPD，对极低的光水平有反应，对低病毒滴度的敏感性将是可能的。 该提案征集资金，以开发一种在快速，自动化的两步分析中检测和识别病毒病毒的工具，该分析可以由急救人员使用，这些急救人员正在响应可疑释放机载（Waterborne）代理。第一步筛选样品，以确定是否有任何具有靶向裸露外套蛋白的病毒颗粒。如果答案是肯定的，则将病毒分离，并分析其核酸以确定其携带病原体基因。