Development of Diagnostic Microarray for Influenza A

描述（由申请人提供）：拟议工作的长期目标是开发一种可用于快速诊断甲型流感的现场便携式微芯片检测方法。甲型流感病毒每年在全球造成50万至100万人死亡，并在感染期间造成直接和间接的生产力损失，对人类造成重大影响。由于甲型流感是一种容易传播的，主要是空气传播的病原体，而且由于这种病毒存在被基因改造成新形式的可能性，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)病毒对人类健康的影响和全球监测的重要性；2)甲型流感作为生物恐怖主义制剂的潜力；3)这种病毒可以作为开发和彻底测试快速、现场便携式诊断的一般方法和技术的极好模型系统。我们将利用功能基因组学的优势，开发一种DNA微阵列，即“流感芯片”，它将提供有关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，这种流感芯片可以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，本工作所开发的技术可以迅速扩展到同时筛选多种生物病原体。该应用程序的具体目标是：1)为甲型流感的分型、分型和菌株分析选择合适的基因靶点；2)优化样品处理和微阵列上的RNA捕获；3)开发芯片上信号放大的新方法；4)开发现场便携式微阵列读取器；5)独立评估和验证流感芯片。为了实现我们的目标，我们召集了一个由学术界（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).