Advanced Microarray Technology for Pathogen Surveillance

描述（由申请人提供）：本提案是对NIAID生物防御合作研究伙伴关系（RFAAI-05-019）的响应。与本RFA的既定目标一致，我们由来自商业部门、学术界和政府的化学家、生物学家和工程师组成的团队建议开展必要的研究，以推进三种潜在的产品，这些产品有望显著提高微阵列在病原体监测和生物防御方面的效用。流感监测将成为该技术的第一个具体应用。由于人类适应型流感病毒是一种容易传播的主要通过空气传播的病原体，并且由于该病毒存在被基因改造成新形式的可能性，甲型流感是一个严重的公共卫生和生物防御问题。疾病控制和预防中心和世界卫生组织强烈强调全球流感监测的重要性，因为它在确定病毒株鉴定（疫苗生产所需）和疾病分布方面发挥着关键作用。虽然有83个国家的112个机构作为国家流感监测中心，但对世界人口的覆盖范围仍然相当有限，需要作出重大努力来加强流感监测。这种缺乏覆盖的一个主要因素是当前昂贵和劳动密集（天）的应变分析方法。为了实现微阵列平台在快速疾病诊断和病原体鉴定方面的巨大潜力，有必要开发替代的、廉价的和易于使用的检测技术。在生物防御中使用微阵列作为有效的监测系统还需要现场便携性，以便迅速和广泛地部署。我们的愿景是开发一种诊断试剂盒，使微阵列的现场应用能够在数小时内而不是数天内进行分析，并且成本与全球分布的需求一致。该试剂盒将提供从样品中快速提取目标的手段，芯片上的信号放大产生肉眼可检测的聚合物，以及用于解释多病原体微阵列的手持式读取器。提出的具体目标是开发以下三种产品：a)用于病毒预浓缩的简单样品处理试剂盒，b)用于直接在微阵列上放大生物识别事件（包括核酸和蛋白质）信号的创新非酶技术，以及c)强大，廉价的现场便携式微阵列读取器。

DESCRIPTION (provided by applicant): This proposal is in response to NIAID's call for Cooperative Research Partnerships for Biodefense (RFAAI-05-019). Consistent with the stated objectives of this RFA, our established team of chemists, biologists and engineers from the commercial sector, academia and government, proposes to conduct the research necessary to advance three potential products that are anticipated to significantly enhance the utility of microarrays for pathogen surveillance and biodefense. Influenza surveillance will be targeted as the first specific application of the technology. Because human-adapted influenza virus is a readily transmitted, primarily airborne pathogen, and because the potential exists for the virus to be genetically engineered into novel forms, influenza A represents a serious public health and biodefense concern. The Centers for Disease Control and Prevention and the World Health Organization have strongly emphasized the importance of global influenza surveillance because of its critical role in determination of viral strain identification (required for vaccine production) and disease distribution. While there are 112 institutions in 83 countries that serve as National Influenza Centers for surveillance, coverage of the world's population remains quite limited, and significant efforts are needed to enhance influenza surveillance. A major factor in this dearth of coverage is the current expensive and labor-intensive (days) method for strain analysis. In order to realize the tremendous potential of the microarray platform for rapid disease diagnosis and pathogen identification, it is essential that alternative, inexpensive and easy-to-use detection technologies be developed. Use of microarrays as effective monitoring systems in biodefense also requires field portability for prompt and widespread deployment. Our vision is a diagnostic kit that would enable field application of microarrays with an analysis time of hours instead of days and at a cost consistent with the need for world-wide distribution. The kit would provide the means for rapid target extraction from the sample, on-chip signal amplification resulting in a polymer detectable by eye, and a hand-held reader to be used for interpretation of multi-pathogen microarrays. The proposed Specific Aims are to develop the following three products: a) a simple sample handling kit for virus pre-concentration, b) an innovative non-enzymatic technology for amplifying signals from bio-recognition events (both nucleic acids and proteins) directly on microarrays, and c) a robust, inexpensive field-portable microarray reader.