Detection and Characterization of Flavivirus and Alphavirus Infections in Acute Suspected Arboviral Cases or Neurological Disease, Central Department, Paraguay

急性疑似虫媒病毒病例或神经系统疾病中黄病毒和甲病毒感染的检测和表征，巴拉圭中央部门

• 批准号: 9808957
• 负责人: Jesse Waggoner
• 金额: $ 23.51万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-12 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9808957
• 关键词: Accounting; Acute; Advanced Development; Alphavirus; Alphavirus Infections; Antibodies; Antibody Response; Arbovirus Infections; Arboviruses; Biological Assay; CRISPR/Cas technology; Categories; Chikungunya virus; Clinical; Clinical Management; Data; Dengue; Dengue Virus; Detection; Diagnosis; Disease; Disease Outbreaks; Encephalitis; Epidemic; Epidemiology; Exposure to; Family; Fever; Flavivirus; Flavivirus Infections; Goals; Guillain-Barré Syndrome; Heterophile Antibodies; Human; Immunity; Immunoglobulin G; Immunoglobulin M; Incidence; Infection; Intervention; Laboratories; Laboratory Diagnosis; Meningitis; Meningoencephalitis; Methods; Molecular; Neurologic; Neurologic Process; Outcome; Pan Genus; Paraguay; Patient risk; Patient-Focused Outcomes; Patients; Phylogenetic Analysis; Population; Protocols documentation; Research; Risk; Risk stratification; Sampling; Serologic tests; Serological; Seroprevalences; Surveillance Methods; Syndrome; System; Techniques; Technology; Test Result; Testing; Viral; Viral Epidemiology; Viral Genome; Viral Load result; Virus; Virus Diseases; West Nile virus; Work; Zika Virus; arbovirus disease; base; chikungunya; clinical Diagnosis; cost; genetic signature; genome sequencing; human disease; human pathogen; improved; nervous system disorder; next generation sequencing; novel; pathogen; patient population; prognostic assays; transmission process; whole genome

PROJECT ABSTRACT Flaviviruses and alphaviruses are the two most common genera of arboviral pathogens, accounting for hundreds of millions of infections annually and repeated epidemics over the past 20 years. In total, these large genera include over a hundred potential human pathogens. Symptomatic infections with the flaviviruses (e.g. dengue, Zika, West Nile) and alphaviruses (e.g. chikungunya, Mayaro) present with non-specific manifestations and predominantly result in two overlapping acute clinical syndromes: a systemic illness with or without fever and a neurological illness such as meningoencephalitis. However, it is costly and impractical to test patients for more than a few common pathogens, and research protocols typically focus on one clinical syndrome. These factors complicate the detection of arboviral infections, surveillance and control efforts, and the calculation of accurate incidence estimates. Research in this proposal will utilize novel molecular methods for pan-genus detection and new multiplex serological methods to simultaneously screen for antibodies against eight arboviruses. The objective of the proposal is to evaluate new molecular and serological methods for the characterization of the incidence, seroprevalence and diversity of flavivirus and alphavirus infections among symptomatic patients. Research will be performed in Asunción and the Central Department of Paraguay, where recent transmission of multiple arboviruses has been documented. Our central hypotheses are that 1) these new methods will increase the breadth of arbovirus detection among symptomatic patients, 2) evidence of exposure to multiple arboviruses will be common, and 3) broad, heterotypic antibody responses will protect against severe disease. This research will first characterize flavivirus and alphavirus infections among patients with an acute suspected arboviral illness or acute neurological disease (meningitis, encephalitis, Guillain-Barré syndrome). Pan-flavivirus and pan-alphavirus detection will be performed with a novel molecular assay. Viral epidemiology, clinical findings, and patient outcomes will be described among patients who meet the study definitions. The incidence of symptomatic and severe disease will then be determined in relation to arboviral antibodies at presentation. Multiplex serological testing will be performed to detect IgM and IgG against five flaviviruses and three alphaviruses. Seroprevalence will be calculated, and the incidence of symptomatic and severe disease will be evaluated in relation to the presence of pre-infection antibodies to heterotypic viruses. Finally, whole-genome viral sequences will be obtained directly from clinical samples using a CRISPR/CAS9 depletion system to analyze viral phylogenetics and identify new/rare viral strains. The purpose of this research is to improve laboratory methods for the most common human arbovirus families and identify patients at risk for symptomatic or severe arboviral illnesses. This work is expected to have an important positive impact because improved surveillance methods and patient risk stratification will result in increased arbovirus detection and targeted interventions for clinical management and outbreak control.

项目摘要 黄病毒属和甲病毒属是虫媒病毒病原体中最常见的两个属， 在过去20年里，每年有数亿人感染，而且一再发生流行病。总的来说，这些大型 属包括超过一百种潜在的人类病原体。黄病毒的症状感染（例如， 登革热、寨卡病毒、西尼罗河病毒）和甲病毒（如基孔肯雅病毒、马亚罗病毒）， 临床表现和主要导致两个重叠的急性临床综合征：全身性疾病，或 没有发烧和神经系统疾病，如脑膜脑炎。然而，这是昂贵和不切实际的， 测试患者的几种常见病原体，研究方案通常集中在一个临床 综合征这些因素使虫媒病毒感染的检测、监测和控制工作复杂化， 准确估计发病率的计算。这项提案的研究将利用新的分子方法 用于全属检测和新的多重血清学方法，以同时筛选抗 八种虫媒病毒。该提案的目的是评估新的分子和血清学方法， 黄病毒和甲病毒感染的发病率、血清阳性率和多样性的特征， 有症状的病人研究将在亚松西翁和巴拉圭中央省进行， 在那里最近有多种虫媒病毒的传播记录。我们的主要假设是：（1） 这些新方法将增加虫媒病毒在有症状患者中检测的广度，2）证据 暴露于多种虫媒病毒将是常见的，3）广泛的，异型抗体反应将保护 对抗严重的疾病。这项研究将首先描述患者中黄病毒和甲病毒感染的特征 患有急性疑似虫媒病毒疾病或急性神经系统疾病（脑膜炎、脑炎、格林-巴利综合征） 综合征）。将采用一种新型分子检测方法进行泛黄病毒和泛甲病毒检测。病毒 将描述符合研究要求的患者的流行病学、临床结果和患者结局 定义.然后将确定与虫媒病毒感染相关的症状性和严重疾病的发生率。 抗体介绍将进行多重血清学检测，以检测IgM和IgG， 黄病毒和三种甲病毒。将计算血清阳性率，并将有症状和 严重的疾病将根据感染前是否存在针对异型病毒的抗体进行评估。 最后，全基因组病毒序列将使用CRISPR/CAS 9直接从临床样品中获得。 使用病毒耗竭系统分析病毒基因组并鉴定新的/罕见的病毒株。本研究的目的 是改善最常见的人类虫媒病毒家族的实验室方法， 用于有症状或严重的虫媒病毒疾病。预计这项工作将产生重要的积极影响 因为改进的监测方法和患者风险分层将导致虫媒病毒 检测和有针对性的干预措施，用于临床管理和疫情控制。