Development of Near Real-Time, Multiplexed Diagnostics for Viral Hemorrhagic Feve

描述（由申请方提供）：出血热病毒（如埃博拉病毒、马尔堡病毒和拉沙病毒）是目前非洲地方病的罪魁祸首，由于这些疾病的高致死率和低感染剂量，因此被CDC列为A类生物威胁。由于担心释放武器化的埃博拉病毒、马尔堡病毒或拉沙病毒，简单有效的检测和诊断至关重要。虽然存在用于诊断这些病毒感染的几种现有测定法，但它们涉及重要的生物安全考虑，因为测定法不是封闭系统样品-应答系统。这降低了这些检测试剂盒在常规临床检测和护理点环境中使用的能力。我们建议研究纳米光子学技术作为快速和多重检测系统来诊断出血热病毒感染的潜力。我们选择了由波士顿大学研究人员开创的两种技术，光子纳米孔阵列和干涉反射成像作为我们的主要检测平台。我们最初将以竞争的方式开发这两种技术。这两种技术在显示不同抗原和病原体的多重检测的能力方面都显示出了希望。根据灵敏度（<104 PFu/ml）和特异性的具体标准，我们将选择最有前途的技术（或互补组合）来开发集成的样本到答案原型检测器。原型检测器将是一个集成系统，将结合微流体，一个多路检测器，有能力区分埃博拉，马尔堡和拉沙感染。该系统将被设计为从血清开始诊断，并提供一个封闭的系统，样本到答案的诊断，是快速和易于使用。该系统将作为概念验证系统，推动光子技术作为便携式诊断的发展。光子学系统具有快速、无标签系统的优点，具有小型化和廉价制造的既定记录。因此，这些技术为新的便携式设备提供了重要的探索途径。为了实现这些目标，我们组建了一个具有互补专业知识的多学科团队。为了促进检测器技术本身的发展，该团队包括微流体，纳米孔阵列开发和干涉检测方面的专家。为了推动测试和捕获探针开发技术，该团队包括假型开发专家，以允许BSL 2测试和动物模型专家熟悉所有将要分析的VHF病毒。原型将通过与Becton Dickinson合作来促进，Becton Dickinson是一家在病毒诊断开发方面具有丰富经验的公司。基于我们组建的团队的实力，我们相信我们能够很好地研究纳米光子学系统作为低功耗诊断的潜力，这些系统简单，并且具有直接的小型化和应用途径在资源有限的临床和护理点环境中。

DESCRIPTION (provided by applicant): Hemorrhagic fever viruses such as Ebola, Marburg and Lassa are responsible for current endemic diseases in Africa and are classified as Category A biothreats by the CDC because of the high fatality that can be associated with these diseases and their low infectious dose. Because of the concern of the release of weaponized Ebola, Marburg or Lassa, simple and effective detection and diagnostics are essential. While there are several existing assays for diagnosing infection with these viruses, they involve significant biosafety considerations, as the assays are not closed system sample-to-answer systems. This reduces the ability of these assays to be used in routine clinical testing and point-of-care settings. We propose to investigate the potential for nanophotonics technologies as rapid and multiplexed detection systems to diagnose infection with hemorrhagic fever viruses. We have chosen two technologies, pioneered by Boston University researchers, photonic nanohole arrays and interferometric reflectance imaging as our primary detection platforms. We will develop both technologies initially in a competitive manner. Both technologies have shown promise in their ability to show multiplexed detection of different antigens and pathogens. Based on the specific criteria of sensitivity (<104 PFu/ml) and specificity, we will select the most promising technology (or a complementary combination) for development of an integrated sample-to-answer prototype detector. The prototype detector will be an integrated system that will incorporate microfluidics, a multiplexed detector with the capacity to distinguish Ebola, Marburg, and Lassa infection. The system will be designed to initiate diagnosis from serum, and provide a closed-system sample-to-answer diagnostic that is rapid and easy to use. This system will serve as a proof of concept system to drive the development of photonic technologies as portable diagnostics. Photonics systems have the advantage of being rapid, label-free systems with an established record of miniaturization and inexpensive manufacture. Thus, these technologies provide an important avenue of exploration for new portable devices. To accomplish these goals we have assembled a multidisciplinary team with complementary expertise. To facilitate the development of the detector technology itself, the team includes experts in microfluidics, nanohole array development, and interferometric detection. To drive the testing and capture probe development technology, the team includes experts in pseudotype development to allow BSL2-testing and animal model experts familiar with all of the VHF viruses that will be analyzed. Prototyping will be facilitated through collaboration with Becton Dickinson, a company with significant experience in the development of virus diagnostics. Based on the strength of the team that we have assembled, we believe that we are well positioned to properly investigate the potential for nanophotonics systems as low power diagnostics that are simple and have a straightforward path to miniaturization and application in both clinical and point-of-care in resource limited settings.