Multiplex serological assays to support arbovirus diagnosis, surveillance and vaccines

多重血清学检测支持虫媒病毒诊断、监测和疫苗开发

• 批准号: 10162498
• 负责人: Aravinda M. DeSilva
• 金额: $ 41.36万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-11 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10162498
• 关键词: Acute; Aedes; Africa; Alphavirus; Americas; Antibodies; Antibody Response; Antigens; Arbovirus Infections; Arboviruses; Area; Arthropods; Asia; Biological Assay; Blood; Central America; Chikungunya virus; Child; Clinical Research; Clinical Trials; Cohort Studies; Consumption; Culicidae; Dengue; Dengue Infection; Dengue Vaccine; Dengue Virus; Detection; Development; Diagnosis; Disease; Drops; Ecology; Environment; Epidemic; Epitopes; Exposure to; Far East; Fever; Flavivirus; Flavivirus Infections; Fluorescence; Human; Immune; Immunity; Individual; Infection; Laboratories; Latin America; Length; Location; Mayaro virus; Measures; Methods; Microspheres; Monitor; North Carolina; Performance; Persons; Phase III Clinical Trials; Population; Proteins; Recombinants; Recording of previous events; Risk; Safety; Sampling; Serology; Serology test; Serotyping; Serum; Specificity; Specimen; Testing; Ticks; Time; Undifferentiated; United States National Institutes of Health; Vaccines; Viral Proteins; Virion; Virus; Yellow fever virus; Zika Virus; Zika virus vaccine; accurate diagnosis; base; cross reactivity; detection assay; experience; feeding; minimally invasive; multiplex assay; neutralizing antibody; novel; predicting response; programs; sample collection; severe dengue; urban area; urban setting; vaccine candidate; vaccine efficacy; vaccine safety; vaccine trial; vaccine-induced antibodies

Abstract At a global level, ongoing ecological, environmental and demographic changes favor the survival and expansion of several mosquito and tick species that transmit arboviruses. Aedes mosquito species that thrive in urban environments created by humans are responsible for epidemics of several flaviviruses [dengue virus (DENV) serotypes 1, 2, 3, 4; Zika virus (ZIKV); yellow fever virus (YFV)] and alphaviruses [chikungunya virus (CHIKV) and Mayaro virus (MAYV)]. Laboratory-based diagnosis and surveillance for arboviruses is difficult because most infected individuals are asymptomatic or develop a mild undifferentiated febrile illness. Serological assays have been developed for the detection of recent or past arboviral infections, but the utility of these assays is severely limited by antibody cross reactivity between related viruses. For example, when ZIKV emerged in many regions of the Americas where greater than 80% of the population was dengue-immune, with current serological assays, it was difficult, if not impossible, to identify infected individuals or monitor the spread of ZIKV at a population level, and likewise to now detect new DENV infections in areas that experienced intense ZIKV epidemics. Our studies over the past 10 years demonstrate that people exposed to flavivirus infections reliably develop antibodies to epitopes that are unique to each flavivirus as well as cross-reactive antibodies. Using our discoveries about the location of immunodominant virus type-specific epitopes, we have produced novel recombinant antigens and demonstrated their utility for the type-specific diagnosis of arboviruses. Under Specific Aim 1 of this proposal, we will build on these discoveries to develop a sample-sparing, microsphere bead-based multiplex assay for the type-specific and sensitive detection of recent or past arbovirus infections. Our initial studies will focus on 8 arboviruses transmitted by Aedes aegypti and Aedes albopictus mosquitos because these viruses share a similar ecology and co-circulate in the same human populations. At a second stage, we will expand the coverage of the assay to detect infections with other arboviruses transmitted by other mosquito species and ticks. Several tetravalent DENV and ZIKV vaccines are currently being evaluated in human clinical trials. While vaccine developers have relied on neutralizing antibodies as a correlate of protection, recent results from clinical trials demonstrate that neutralizing antibodies alone are a poor correlate of vaccine safety and efficacy. We have identified flavivirus type- and epitope-specific antibody responses that are better predictors than neutralizing antibodies of vaccine safety and efficacy. Under Specific aim 2 of this proposal, we will develop a sample-sparing microsphere-based assay for the detection of epitope-specific vaccine-induced antibody responses that are correlated with protective immunity to each of the DENV serotypes and ZIKV. The technological advances and products from this proposal will enhance our ability to efficiently monitor arbovirus infections at the individual and population levels and also support the development of arbovirus vaccines.

抽象的 在全球范围内，正在进行的生态，环境和人口变化有利于生存和扩张 在传播arbovirus的几种蚊子和壁虱物种中。在城市中繁衍生息的艾德斯蚊子 人类创造的环境负责几种黄病毒的流行病[登革热病毒（DENV） 血清型1、2、3、4;寨卡病毒（Zikv）;黄热病病毒（YFV）]和α病毒[Chikungunya病毒（CHIKV） 和Mayaro病毒（Mayv）]。基于实验室的Arboviruses诊断和监视很困难，因为 大多数受感染的人无症状或患有轻度的未分化的高温疾病。血清学测定 已经开发用于检测近期或过去的弧病毒感染，但这些测定的实用性是 受相关病毒之间的抗体交叉反应性严重限制。例如，当Zikv出现在许多 美洲地区大于80％的人口是登革热的，目前的血清学 测定，很难（即使不是不可能）识别受感染的个体或监测ZIKV的传播 人口水平，同样可以在经历强烈ZIKV的地区发现新的DENV感染 流行病。过去十年来我们的研究表明，暴露于黄病毒感染的人们可靠 开发针对每种黄病毒和交叉反应抗体独有的表位的抗体。使用我们的 关于免疫主导病毒类型特异性表位的位置的发现，我们已经产生了新颖的 重组抗原，并证明了它们用于特定于arboviruses类型诊断的实用性。在下面 该提案的具体目的1，我们将基于这些发现，以开发出样本比例， 基于微球珠的多重分析，用于针对最近或过去的类型特异性检测 arbovirus感染。我们的最初研究将重点放在埃及埃及和艾德斯传输的8种Arbovires上 白化蚊子因为这些病毒具有相似的生态学并在同一人类中共同循环 人群。在第二阶段，我们将扩大测定的覆盖范围，以检测与其他的感染 其他蚊子和壁虱传播的arbovirus。几种四位化的DENV和ZIKV疫苗是 目前正在人类临床试验中进行评估。疫苗开发人员依靠中和 抗体作为保护的相关性，临床试验的最新结果表明，中和抗体 仅疫苗安全性和功效的相关性很差。我们已经确定了黄病毒类型和表位特异性 抗体反应比中和疫苗安全性和功效的抗体更好。在下面 该提案的具体目的2，我们将开发出基于样本的微球测定法 与保护性相关的表位特异性疫苗诱导的抗体反应的检测 对每种DENV血清型和ZIKV的免疫力。从中的技术进步和产品 提案将增强我们在个人和人群水平上有效监测Arbovirus感染的能力 并支持arbovirus疫苗的开发。