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 黄病毒和甲病毒是虫媒病毒病原体中最常见的两个属，占 过去 20 年来，每年有数亿人感染并反复流行。 属包括一百多种潜在的人类病原体黄病毒感染（例如黄病毒）。 登革热、寨卡病毒、西尼罗河病毒）和甲病毒（例如基孔肯雅病毒、马亚罗病毒）存在非特异性 表现并主要导致两种重叠的急性临床综合征：一种全身性疾病，伴有或 没有发烧和神经系统疾病，例如脑膜脑炎，但是成本高昂且不切实际。 对患者进行多种常见病原体测试，研究方案通常集中于一种临床 这些因素使虫媒病毒感染的检测、监测和控制工作变得复杂化。 该提案中的研究将利用新颖的分子方法来计算准确的发病率估计。 用于泛属检测和新的多重血清学方法，以同时筛选抗 该提案的目的是评估八种虫媒病毒的新分子和血清学方法。 黄病毒和甲病毒感染的发病率、血清流行率和多样性的特征 研究将在亚松森和巴拉圭中部地区进行 最近记录了多种虫媒病毒的传播，我们的中心假设是 1)。 这些新方法将扩大有症状患者中虫媒病毒检测的广度，2）证据 接触多种虫媒病毒将很常见，并且 3) 广泛的异型抗体反应将保护 这项研究将首先确定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、吉兰-巴雷病） 泛黄病毒和泛甲病毒检测将通过新型分子病毒检测进行。 将在符合研究的患者中描述流行病学、临床发现和患者结果 然后将根据虫媒病毒确定有症状和严重疾病的发生率。 提交时将进行多重血清学检测，以检测针对 5 种抗体的 IgM 和 IgG。 将计算黄病毒和三种甲病毒的血清阳性率，以及症状和症状的发生率。 严重疾病将根据感染前异型病毒抗体的存在进行评估。 最后，将使用 CRISPR/CAS9 直接从临床样本中获得全基因组病毒序列 耗竭系统分析病毒系统发育并鉴定新的/稀有的病毒株本研究的目的。 旨在改进最常见人类虫媒病毒家族的实验室方法并识别处于危险中的患者 这项工作预计会对有症状或严重的虫媒病毒疾病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增加 临床管理和疫情控制的检测和有针对性的干预措施。