Detection and discovery of viral pathogens associated with dengue-like symptoms

大规模的病毒监测研究对于监测全球传染病的出现至关重要。 疾病和产生关于流行病毒种类和季节性的重要信息 新物种的可能存在。在中国进行的许多大型流行病学研究 然而，温带国家没有在热带环境中重复出现，很可能会暴露出病毒负担。 以及由于固有的环境和社会差异而导致的发病率变化。登革热病毒是一种 一种典型的新发传染病，每年迅速感染5000-1亿人 地理范围扩大，是人类最常见的媒介传播病毒疾病。目前 然而，现有的实验室诊断方法无法检测出30%-70%的出现DV感染的患者 有明显的登革热症状。这项研究的目的有两个：利用现有的最佳诊断方法 在发展中的热带环境中描绘病毒病原体光谱的技术 利用广泛筛查提供的数据作为平台，以便 发现新的或不同的人类病毒病原体。 在与伊娃·哈里斯的合作下，这项研究将使用病毒芯片(UCSF)来表征人类血清 和来自尼加拉瓜患者的PBMC样本，这些样本来自拟议的基于医院的研究 多个季节出现类似登革热的症状。这将使我们能够捕捉到流行率、季节性 随着时间的推移，所有已知病毒在疑似登革热病例中的双重感染率。我们还建议利用 根据几个标准对选定数量的样本进行超深度测序，包括严重程度 发现以前没有特征的病原体的症状。我们将进行全基因组克隆和 有希望的线索的分子表征，包括基于聚合酶链式反应的快速诊断和 用于后续相关性研究的血清学试剂。

Large-scale viral surveillance studies are critical to monitoring the global emergence of infectious disease and yield important information about the prevalence and seasonality of circulating virus species and the possible existence of novel species. Many large epidemiologic studies that have been conducted in temperate countries have not been repeated in tropical settings, however, and are likely to reveal viral burden and incidence variations due to inherent environmental and social differences. Dengue virus (DV) is an archetypal emerging infectious disease that infects an estimated 50-100 million people annually in a rapidly expanding geographic range and is the most common vector-borne viral disease of humans. Currently available lab diagnostic methods, however, fail to detect DV infection in 30-70% of patients presenting with hallmark dengue symptoms. The purpose of this study is twofold: to use the best available diagnostic technologies to delineate the spectrum of viral pathogens in a developing tropical environment with respect to dengue-like illnesses and to utilize the data provided by the extensive screening as a platform for the discovery of novel or divergent human viral pathogens. In collaboration with Eva Harris, this study will use the Virochip (UCSF) to characterize human sera and PBMC samples from Nicaraguan patients from the proposed hospital-based study presenting with dengue-like symptoms over multiple seasons. This will allow us to capture both the prevalence, seasonality and dual infection rates for all known viruses in suspected dengue cases over time. We also propose to utilize ultra-deep sequencing for a select number of samples based on several criteria including severity of symptoms for discovery of previously uncharacterized agents. We will pursue full genome cloning and molecular characterization for promising leads, including development of PCR-based rapid diagnostics and serological reagents for subsequent association studies.