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

病毒性出血热近实时多重诊断的开发

• 批准号: 8184838
• 负责人: John H Connor
• 金额: $ 142.23万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8184838
• 关键词: Africa; Animal Model; Antigens; Biological Assay; Biosensor; Boston; Categories; Centers for Disease Control and Prevention (U.S.); Clinical; Collaborations; Communicable Diseases; Communities; Detection; Development; Devices; Diagnosis; Diagnostic; Diagnostic tests; Differential Diagnosis; Discipline; Disease; Disease Outbreaks; Dose; Ebola Hemorrhagic Fever; Endemic Diseases; Engineering; Epidemic; Etiology; Generations; Goals; Image; Individual; Infection; Infectious Agent; Label; Light; Medical; Methods; Microfluidics; Miniaturization; Nucleic Acid Amplification Tests; Particle Size; Pathogen detection; Patients; Positioning Attribute; Preparation; Process; RNA Viruses; Reporting; Research Personnel; Resources; Sampling; Satellite Viruses; Serum; Small RNA; Specificity; Symptoms; System; Techniques; Technology; Telecommunications; Testing; Time; Universities; Viral; Viral Hemorrhagic Fevers; Virion; Virus; Work; base; biodefense; biothreat; clinically relevant; design; detector; experience; flu; hemorrhagic fever virus; large scale production; multidisciplinary; nanopattern; next generation; pathogen; photonics; point of care; point-of-care diagnostics; prototype; rapid detection; research clinical testing; response; sensor; success; technology development; tool; virus development

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. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to develop a proof-of-concept design for a self-contained diagnostic system for viruses that cause hemorrhagic fever. The basis for the diagnostic platform will be a light-based detector that will allow low-power, simple testing of patients that are potentially infected. The straightforward technology that is proposed here can be directly scaled to inexpensive large-scale production through the leveraging of existing telecommunications and computing manufacturing techniques.

描述（由申请人提供）：埃博拉病毒、马尔堡病毒和拉沙病毒等出血热病毒是导致非洲当前地方性疾病的原因，由于与这些疾病相关的高致死率和低感染剂量，它们被疾病预防控制中心列为A类生物威胁。由于担心武器化的埃博拉病毒、马尔堡病毒或拉沙病毒的释放，简单有效的检测和诊断至关重要。虽然已有几种诊断这些病毒感染的检测方法，但它们涉及重大的生物安全考虑，因为这些检测方法不是封闭系统，从样本到答案的系统。这降低了这些检测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路检测系统诊断出血热病毒感染的潜力。我们选择了两种由波士顿大学研究人员首创的技术，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。我们将首先以竞争的方式开发这两项技术。这两种技术都显示出对不同抗原和病原体进行多重检测的能力。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）来开发集成的样品到答案原型检测器。原型检测器将是一个集成系统，将包括微流体，一个具有区分埃博拉，马尔堡和拉沙感染能力的多路检测器。该系统将被设计用于从血清开始诊断，并提供一种快速且易于使用的从样本到答案的封闭系统诊断。该系统将作为概念验证系统，推动便携式光子诊断技术的发展。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，这些技术为探索新的便携式设备提供了一条重要的途径。为了实现这些目标，我们组建了一支具有互补专业知识的多学科团队。为了促进探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，该团队包括允许bsl2测试的伪型开发专家和熟悉将要分析的所有甚高频病毒的动物模型专家。将通过与在病毒诊断开发方面具有丰富经验的Becton Dickinson公司合作，促进原型制作。基于我们所组建的团队的实力，我们相信我们已经做好了充分的准备，可以适当地研究纳米光子学系统作为低功耗诊断的潜力，这种诊断简单，并且在资源有限的情况下，可以直接实现小型化，并应用于临床和护理点。