A Rapid and Specific Diagnostic for Immunoglobulin Response to Zika Virus Exposure Based on De novo Designed Low Cost Hyper-stable Mini-Protein Epitopes

项目摘要 寨卡病毒是一种由蚊子传播的病原体， 感染妇女所怀胎儿的并发症。最近，寨卡病毒与一个 格林-巴利综合征和小头畸形病例数量激增， 婴儿的大脑发育受到严重阻碍，出生时头部较小。鉴于这种 可能与这些严重的出生缺陷有关，迫切需要一种方法来确定 诊断一名妇女是否接触过病毒。目前的测试方法 寨卡病毒的存在错过了感染后病毒清除身体之前的狭窄时间窗口 和血清学试验往往受到交叉反应，以其他黄病毒流行的， 与寨卡病毒相同的地区。这里描述的研究提出了一种 免疫测定法仅靶向宿主多克隆应答中的那些抗体， 寨卡病毒外壳蛋白中不被其他黄病毒共享的表位。这样的测定 可能会导致更多的临床特异性免疫检测的病毒问题。首先我们将 开发新的计算机设计的寨卡病毒超稳定病毒表位模拟物（VEM） 具有可特异性结合人抗寨卡免疫球蛋白的结构的包膜蛋白。 第二，这些VEM将以快速、低成本的纸基三明治的形式实现 能够在临床相关条件下检测人血浆中的抗寨卡抗体的免疫测定法 浓度的

PROJECT ABSTRACT The Zika virus is a mosquito-borne pathogen that has been implicated in a number of grave complications to fetuses carried by infected women. More recently, Zika has been linked to a spike in the number of cases of Guillain-Barre syndrome and microcephaly, a condition in which babies have severely hindered brain development and are born with smaller heads. Given this possible link to these serious birth defects, there is a pressing need for a way to definitively diagnose whether a woman has been exposed to the virus. Current methods to test for the presence of Zika miss the narrow time window before the virus clears the body after infection and serological tests are often hindered by cross-reactivity to other flaviviruses endemic in the same regions as Zika. The studies described here propose the development of an immunoassay targeting only those antibodies in the host’s polyclonal response that bind to those epitopes in the Zika coat protein that are not shared by other Flaviviruses. Such an assay could lead to more clinically specific immunoassays for the viruses in question. First we will develop novel computationally designed hyperstable viral epitope mimetics (VEMs) of the Zika envelope protein with structures that can specifically bind human anti-Zika immunoglobulins. Second these VEMs will be implemented in a rapid, low-cost paper-based sandwich immunoassay capable of detecting anti-Zika antibodies in human plasma at clinically relevant concentrations.