Structure based design of recombinant Zika virus antigens for serodiagnosis

摘要 这项建议的重点是合理设计用于寨卡病毒(ZIKV)血清诊断的抗原，并将其用于最低限度的 与其他人类致病性黄病毒的交叉反应。寨卡病毒(ZIKV)感染会导致流产， 小头畸形和格林-巴利综合征。迫切需要对ZIKV感染进行特异性血清学诊断 跟踪ZIKV感染的传播，并在产前等情况下诊断暴露于ZIKV的个人 诊断和新生儿监测、献血者筛查、寨卡疫情归国旅行者筛查 地区。寨卡病毒血清诊断具有很高的挑战性，因为它与登革热和其他病毒具有广泛的交叉反应 与寨卡病毒共同传播的黄病毒。我们之前和正在进行的定义人类血清型特异性的工作 登革热包膜(E)蛋白的抗体反应使我们处于独特的位置来定义相似的区域 ZIKV E，并产生用于血清诊断的候选诊断抗原。用详细的结构加固 信息，我们将合理设计和生产具有独特抗原性的寨卡病毒E蛋白亚结构 黄病毒初免、免疫前黄病毒感染的特异性和敏感性血清学检测 或接种了黄病毒疫苗的人群。对于目标1，我们将生产ZIKV候选抗原并测量它们的 暴露于初级或慢性接触者的免疫血清参考板上的抗体结合反应 继发性黄病毒感染，包括ZIKV和DENV。在目标2中，我们将使用蛋白质工程 重新设计最有效的候选抗原以改变交叉反应抗原部位的方法，而 保留寨卡病毒特异性位点，提高抗原对寨卡病毒血清学鉴别诊断的潜力 寨卡病毒感染。我们将对一种或一种表面重新设计的寨卡病毒抗原进行比较和对比 最近和以前接触寨卡病毒的血清学检测。在后续研究中，我们将与参考资料合作 实验室和商业伙伴使用重组蛋白抗原开发简单的分析方法，用于 看护的重点和监控项目。

Abstract This proposal focuses on the rational design of antigens for serodiagnosis of Zika virus (ZIKV) with minimal cross-reactivity to other human pathogenic flaviviruses. Zika virus (ZIKV) infection can cause miscarriage, microcephaly, and Guillain-Barré syndrome. A specific serodiagnosis of ZIKV infection is urgently needed to track the spread of ZIKV infection and to diagnose individuals exposed to ZIKV in scenarios such as prenatal diagnosis and neonatal monitoring, blood donor screening, screening of travelers returning from Zika-endemic regions. ZIKV serodiagnosis is highly challenging because of extensive cross-reactivity with dengue and other flaviviruses that co-circulate with ZIKV. Our previous and ongoing work defining the human serotype specific antibody responses to dengue envelope (E) protein place us in a unique position to define similar regions in ZIKV E and produce candidate diagnostic antigens for serodiagnostics. Fortified with detailed structural information, we will rationally design and produce substructures of Zika E protein bearing unique antigenic sites for specific and sensitive serological detection of ZIKV infections in flavivirus naïve, flavivirus pre-immune or flavivirus-vaccinated populations. For aim 1, we will produce ZIKV candidate antigens and measure their antibody binding response across a reference panel of immune sera from people exposed to primary or secondary flavivirus infections, including ZIKV and DENV. In Aim 2, we will use protein engineering approaches to redesign most efficacious candidate antigens to change cross-reactive antigenic sites, while preserving Zika specific sites, to improve the diagnostic potential of antigens for differential serodiagnosis of Zika infection. We will compare and contrast one or a combination of surface redesigned Zika antigens for serological detection of recent and prior exposure to ZIKV. In follow up studies, we will partner with reference laboratories and commercial partners to use recombinant protein antigens to develop simple assays for use at the point of care and in surveillance programs.