Development of a novel test for the early detection of Dengue Virus infection

描述（由申请人提供）：登革热病毒（DENV）是一个日益重要的全球重大公共卫生问题，西半球约有25亿人面临感染风险，每年报告5000 -1亿例病例，每年相关费用为21亿美元[1,2]。登革出血热是一种蚊媒病毒，是登革热、登革出血热和登革休克综合征的病原体，由于这种发热性疾病的非特异性临床表现，登革出血热常被误诊。有超过1700万美国人到流行地区旅行，现在在返回美国的旅行者中，登革热感染比疟疾感染更常见。迫切需要快速、敏感和特定的医疗点兼容的DENV诊断工具，以改善患者护理和公共卫生监测。在本应用中，我们将探索一种新的捕获技术CAP-TECH的发现，该技术专为特异性捕获DENV （ViCAP-DENV）而设计，该技术构建在护理点兼容的固相顺磁微颗粒上，并在荧光免疫分析（FIA）中使用，可以在几分钟内快速检测DENV。此外，在一项比较研究中，ViCAP-DENV FIA原型观察到的荧光信号的灵敏度与qRT-PCR观察到的信号相当，但时间和成本都是前者的一小部分。我们最重要的假设是，使用一种诊断工具，如ViCAP-DENV FIA，将能够快速和敏感地检测血液样本中的登革热病毒，最终使用该工具可以改善患者的预后。进一步优化ViCAP-DENV和FIA（目标1），并对ViCAP-DENV FIA性能进行初步临床评估，与fda批准的DENV测试方法进行对比研究，使用可持续科学研究所和尼加拉瓜卫生部收集的去鉴定血清样本（目标2），将为未来的产品开发和II期现场测试评估提供基础工作。该应用程序的成功将提供一种新颖且具有成本效益的护理点兼容测试设计，可在几分钟内从样本中快速诊断病毒感染。

DESCRIPTION (provided by applicant): Dengue virus (DENV) is a major public health problem of growing global importance with ~2.5 billion people at risk of infection, 50-100 million cases reported annually and associated costs of $2.1 billion per year in the Western Hemisphere [1,2]. DENV is a mosquito-borne virus and the causative agent of dengue fever, dengue hemorrhagic fever and dengue shock syndrome and is often misdiagnosed due to the non-specific clinical manifestations of this febrile illness [3]. With more than 17 million Americans traveling to endemic areas, dengue infections are now more common in travelers returning to the U.S. than malaria [16]. There is an urgent need for rapid, sensitive and specific point-of-care-compatible DENV diagnostic tools for improved patient care and public health surveillance. In this application, we will explore the discovery that a novel capture technology, CAP-TECH, designed for the specific capture of DENV (ViCAP-DENV) constructed on point-of-care-compatible solid-phase paramagnetic microparticles and their use in a fluorescent immunoassay (FIA) enabled the rapid detection of DENV in minutes. Further, in a comparative study, the sensitivity of fluorescent signal observed by the ViCAP-DENV FIA prototype was comparable to the signal observed by qRT-PCR, but in a fraction of the time and cost. Our overriding hypothesis is that the use of a diagnostic tool, such as ViCAP-DENV FIA, will enable the rapid and sensitive detection of Dengue virus in blood samples, the eventual use of which could improve patient outcomes. Further optimization of ViCAP-DENV and the FIA (Aim 1) and an initial clinical evaluation of ViCAP- DENV FIA performance in a side-by-side comparison study against FDA-approved DENV test methods using de-identified serum samples collected by the Sustainable Sciences Institute and the Nicaraguan Ministry of Health (Aim 2), will provide the foundational work for future product development and field-testing evaluations in Phase II. The success of this application will provide a novel and cost-effective point-of-care-compatible test design for the rapid diagnosis of viral infection from samples within minutes.