Receptor Mimics for Rapid Detection, Typing, and Susceptibility Testing of Influe

描述(申请人提供)：流感造成的疾病负担很大。在撰写这份提案时，2009年H1N1大流行正在展开。截至2009年8月23日，世界卫生组织报告了超过209,438例经实验室确认的2009年H1N1流感病例，至少2,185人死亡。具有测试病毒对当前抗病毒药物敏感性的快速、准确、护理点检测试剂盒在我们抗击这种致命病毒的斗争中至关重要。该方案的目标是开发针对病毒表面丰富的病毒糖蛋白血凝素(HA)和神经氨酸酶(NA)的新型糖蛋白诊断平台(~50拷贝的四聚体NA和300拷贝的三聚体HA)。这一提议的基本原理是，一个设计良好的葡聚糖微阵列平台将在暴露于病毒的情况下产生“指纹识别模式”。这一建议的新颖之处在于，可以快速评估对达菲(R)和瑞乐沙(R)的敏感性，而不需要耗时地识别和表征新出现的菌株。在具体目标1中，我们将建立一个聚糖库，以获取和评估流感的抗病毒敏感性。在具体目标2中，我们将开发不同的诊断平台来捕获和分型流感病毒株。我们还将开发测试，以确定对抗病毒药物的敏感性或耐药性。这些测试尤其重要，因为在确定抗病毒耐药性突变的基因组序列之前，目前还没有快速检测和评估抗病毒敏感性的方法。在具体目标3中，我们将使用临床样本对不同的平台进行评估。具体地说，我们将使用从感染患者中分离的临床样本来检查诊断平台的稳健性、敏感性和选择性，并评估抗病毒敏感性。实现这项提案中的具体目标预计将产生快速、可靠、关注点和用户友好的诊断方法，用于准确检测和区分不同的流感毒株。

DESCRIPTION (provided by applicant): Disease burden due to influenza is significant. The 2009 H1N1 pandemic is unfolding as this proposal is being written. As of August 23, 2009, over 209,438 laboratory-confirmed cases of 2009 H1N1 influenza viruses with at least 2,185 deaths have been reported by the World Health Organization. Rapid, accurate, point-of-care detection kits that possess the capability to test for susceptibility of the virus to current antivirals are critical in our fight against this deadly virus. The objective of this proposal is to develop novel glycan based diagnostic platforms that target the viral glycoproteins, Hemagglutinin (HA) and Neuraminidase (NA), which are abundant (~50 copies of tetrameric NA and 300 copies of trimeric HA) on the surface of the virion. The rationale for this proposal is that a well designed glycan microarray platform will yield a "fingerprint pattern of recognition" on exposure to the virus. The novelty of this proposal is that susceptibility to Tamiflu(r) and Relenza(r), can be rapidly evaluated without the time consuming need to identify and characterize an emerging strain. In Specific Aim 1, we will generate a library of glycans to capture and assess antiviral susceptibility of influenza. In Specific Aim 2, we will develop different diagnostic platforms to capture and type influenza strains. We will also develop tests to determine sensitivity or resistance to antiviral agents. These tests are especially important since there is currently no method to rapidly detect and assess antiviral susceptibility before the genomic sequence for antiviral resistance mutations has been determined. In Specific Aim 3, we will evaluate the different platforms using clinical samples. Specifically, we will examine the robustness, sensitivity, and selectivity of the diagnostic platforms using clinical samples isolated from infected patients and assess antiviral susceptibility. Accomplishing the specific aims in this proposal is expected to yield rapid, robust, point of care, user friendly diagnostics for the precise detection and differentiation of different influenza strains.