Development of Diagnostic Microarray for Influenza A

说明(申请人提供)：拟议工作的长期目标是开发一种现场便携式微芯片分析方法，作为对甲型流感的快速诊断。每年甲型流感病毒感染对人类造成重大影响，包括每年全球50万至100万人死亡，以及感染期间生产力直接和间接损失造成的经济影响。由于甲型流感是一种容易传播的病原体，主要是通过空气传播的，而且这种病毒有可能被基因工程改造成新的形式，因此它代表了一个严重的生物防御问题。我们选择与甲型流感病毒合作有三个重要原因：1)甲型流感病毒对人类健康的影响和全球监测的重要性，2)甲型流感病毒作为生物恐怖主义病原体的潜力，以及3)这种病毒可以作为开发和彻底测试快速现场便携诊断的一般方法和技术的优秀模型系统。我们将利用功能基因组学，开发一种名为“流感芯片”的DNA微阵列，它将提供关于个人是否感染流感的信息，并提供病毒的类型和抗原亚型特征。正如设想的那样，这种流感芯片可能在对基因工程流感病毒的快速防御反应中发挥关键作用。此外，这项工作中开发的技术可以迅速扩展到同时筛查多种生物病原体。这项申请的具体目标是1)为甲型流感的分型、分型和毒株分析选择合适的遗传目标，2)优化样本处理和在微阵列上捕获RNA，3)开发用于芯片信号放大的新方法，4)开发现场便携式微阵列读取器，以及5)对流感芯片进行独立评估和验证。为了实现我们的目标，我们召集了一个由来自学术界(科罗拉多大学)、政府(疾病控制中心)和商业部门(TeleChem/Arrayit.com)的科学家和工程师组成的协调良好的小组。

DESCRIPTION (provided by applicant): The long-term objective of the proposed work is the development of a field portable microchip assay that will serve as a rapid diagnostic for influenza A. Annual influenza A virus infections have a significant impact on humanity both in terms of death, between 500,000 and 1,000,000 people worldwide each year, and economic impact resulting from direct and indirect loss of productivity during infection. Because influenza A is an easily transmitted, primarily airborne pathogen, and because the potential exists for the virus to be genetically engineered into novel forms, it represents a serious biodefense concern. We have chosen to work with influenza A virus for three significant reasons: 1) the virus' impact on human health and the importance of worldwide surveillance, 2) influenza A's potential as a bioterrorism agent, and 3) this virus can serve as an excellent model system for development and thorough testing of general methodologies and technologies for rapid, field portable diagnostics. We will take advantage of functional genomics by developing a DNA microarray, the "Flu Chip," that will provide information as to whether or not an individual is infected with influenza as well as provide both type and antigenic sub-type characterization of the virus. As envisioned, this Flu Chip could play a key role in a rapid defensive response to a genetically engineered influenza virus. In addition, the technology to be developed in this work could be quickly extended to screen for multiple biological pathogens simultaneously. The specific aims of this application are 1) selection of appropriate genetic targets for typing, sub-typing and strain analysis for influenza A, 2) optimization of sample handling and RNA capture onto microarrays, 3) development of new approaches for on-chip signal amplification, 4) development of a field-portable microarray reader, and 5) independent evaluation and validation of the Flu Chip. To accomplish our objectives, we have assembled a well-coordinated group of scientists and engineers from academia (University of Colorado), government (Centers for Disease Control), and the commercial sector (TeleChem/Arrayit.com).