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，每年报告的病例为5000-1亿例，相关费用为每年21亿美元[1，2]。DENV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，经常被误诊[3]。随着1700多万美国人前往流行地区，登革热感染在返回美国的旅行者中比疟疾更常见[16]。迫切需要快速、灵敏和特定的与护理地点兼容的DENV诊断工具，以改善患者护理和公共卫生监测。在这项应用中，我们将探索一项新的捕获技术CAP-TECH，该技术专为特定捕获DENV(ViCAP-DENV)而设计，构建在与护理点兼容的固相顺磁微粒上，并将其用于荧光免疫分析(FIA)，使DENV能够在几分钟内快速检测出来。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，但时间和成本只有一小部分。我们压倒一切的假设是，使用诊断工具，如ViCAP-DENV FIA，将能够快速而灵敏地检测血液样本中的登革病毒，最终使用该工具可能会改善患者的预后。ViCAP-DENV和FIA的进一步优化(AIM 1)，以及对ViCAP-DENV FIA性能的初步临床评估，通过与FDA批准的DENV测试方法进行并列比较研究，使用由可持续科学研究所和尼加拉瓜卫生部收集的未确认的血清样本(AIM 2)，将为未来的产品开发和第二阶段的现场测试评估提供基础工作。这一应用的成功将为在几分钟内从样本中快速诊断病毒感染提供一种新的、具有成本效益的护理点兼容的测试设计。

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.