Development of a Novel, Sensitive ZIKV-Specific Sero-diagnostic Assay Utilizing Biologically Inspired Synthetic Molecules

利用受生物学启发的合成分子开发新型、灵敏的 ZIKV 特异性血清诊断检测方法

• 批准号: 9215750
• 负责人: DONALD SCOTT BURKE
• 金额: $ 69.4万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-18 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9215750
• 关键词: Adult; Affect; Affinity; Antibodies; Antibody Affinity; Antigens; Binding; Biological Assay; Biological Markers; Brazil; Carbon; Child; Clinical; Communicable Diseases; Culicidae; Dengue Virus; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Disease; Disease Outbreaks; Emerging Communicable Diseases; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epitopes; Evaluation; Exanthema; Fever; Flavivirus; French Polynesia; General Population; Generations; Genes; Goals; Grant; Guillain-Barré Syndrome; HIV; HIV Antibodies; HIV Infections; HIV antibody positive; IgG1; IgG2; IgG3; IgG4; Immune; Immunity; Immunoglobulin A; Immunoglobulin M; Immunoglobulins; Incidence; Infection; Investigation; Kinetics; Laboratories; Lead; Libraries; Ligands; Measures; Methodology; Methods; Microcephaly; Micronesia; Molecular; Molecular Bank; Monitor; N-substituted Glycines; Newborn Infant; Nitrogen; Patients; Pattern; Peptides; Peptoids; Performance; Persons; Population; Positioning Attribute; Prevention; Problem Solving; Procedures; Public Health; Reagent; Reporting; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Sensitivity and Specificity; Serum; Side; Staging; Symptoms; Testing; United States National Institutes of Health; Vaccines; Validation; Variant; Vertebral column; Virus Diseases; Work; Zika Virus; base; cohort; combinatorial; cost; cross reactivity; design; diagnostic assay; high throughput screening; human disease; individual patient; innovative technologies; molecular shape; new technology; next generation; novel; novel diagnostics; novel marker; patient biomarkers; programs; response; screening; small molecule; tool

PROJECT SUMMARY/ABSTRACT Zika virus (ZIKV), a mosquito-borne flavivirus genetically related to dengue virus (DENV), produces a mild febrile illness with maculo-papular rash with approximately 80% of patients demonstrating no symptoms. Until 2007 only 14 cases of human disease had been reported since ZIKV's discovery in 1947. In 2015, an epidemic exploded with estimations of over one million current infections in Brazil alone. While ZIKV infections are generally mild, recently they have been associated with Guillain-Barré syndrome in adults and children and an alarmingly high rate of microcephaly in newborns. There are no commercial vaccines or diagnostic assays available. Accurate detection of ZIKV immunity, as measured by anti-ZIKV antibodies, will be a critical tool in prevention and control programs. Current ZIKV sero-diagnostic methods typically show substantial cross- reactivity with other flavivirus reagents. We have recently developed a revolutionary approach to discover novel diagnostic antibody ligands for viral infections (HIV and DENV) by screening a very large combinatorial library (108 molecular variants) of biologically inspired, but non-biological, molecular shapes, or peptoids, against panels of post-infection patient sera. Using this method, we isolated several peptoids that specifically bound antibodies present in post-infection sera that are not present in normal non-immune sera nor pre- infection sera. Our proof-of-concept preliminary studies have demonstrated the ability of peptoid biomarkers to differentiate between HIV antibody positive and HIV antibody negative serum as well as DEN antibody positive and DEN antibody negative serum. Additionally, we have also identified peptoid biomarkers that can distinguish between recent and non-recent HIV infections. We have used these novel non-biological specific ligands as the bases for HIV ELISA assays which display very good sensitivity and specificity. In this grant, we propose to harness this novel technology to develop robust, peptoid-based assays to specifically detect ZIKV antibodies. We will test this innovative technology against panels of sera, assembled from patients in Recife, the epicenter of the Zika outbreak in Brazil. Direct access to clinical cohorts and patient samples positions our team well for a ZIKV diagnostic development. After identification of ZIKV reactive peptoids, we will perform a detailed characterization of the kinetics of the immunoglobulin responses against the selected peptoids. The identification of these peptoid biomarkers will then lead to the development of a second-generation combinatorial library where peptoid reactivity will be further optimized, and we will identify the mechanisms of binding between the ZIKV-specific peptoid biomarkers and the patient antibody. The goals described in this project will lead to the development of a highly optimized, low cost, peptoid-based high-throughput assay to accurately detect ZIKV antibodies and thereby diagnose individual patients and measure incidence in affected populations. The development of such diagnostic assay and disease surveillance tools will greatly increase the capacity of public health agencies worldwide to monitor infectious diseases of global importance.

项目总结/摘要 寨卡病毒（ZIKV）是一种与登革热病毒（DENV）遗传相关的蚊媒黄病毒， 发热性疾病伴斑丘疹，约80%的患者无症状。直到 2007年，自1947年发现ZIKV以来，仅报告了14例人类疾病。2015年，一场流行病 据估计，仅在巴西，目前的感染人数就超过了100万。虽然ZIKV感染是 一般轻度，最近他们与成人和儿童的格林-巴利综合征有关， 新生儿患小头畸形的比例高得惊人。没有商业疫苗或诊断检测 available.如通过抗ZIKV抗体测量的ZIKV免疫的准确检测将是ZIKV免疫的关键工具。 预防和控制方案。目前的ZIKV血清诊断方法通常显示出实质性的交叉免疫。 与其他黄病毒试剂反应。我们最近开发了一种革命性的方法来发现 通过筛选一种非常大的组合的用于病毒感染（HIV和DENV）的新型诊断抗体配体， 生物启发的，但非生物的，分子形状或类肽的文库（108种分子变体）， 针对感染后患者血清组。使用这种方法，我们分离出几种特异性 存在于感染后血清中的结合抗体不存在于正常非免疫血清中，也不存在于感染前血清中。 感染血清我们的概念验证初步研究已经证明了类肽生物标志物的能力， 区分HIV抗体阳性和HIV抗体阴性血清以及DEN抗体阳性 和DEN抗体阴性血清。此外，我们还鉴定了类肽生物标志物， 区分最近和非最近感染艾滋病毒。我们使用了这些新的非生物特异性 配体作为HIV ELISA测定的基础，其显示出非常好的灵敏度和特异性。在这份协议中，我们 我建议利用这种新技术来开发稳健的、基于类肽的检测方法来特异性检测ZIKV 抗体的我们将用从累西腓病人身上收集的血清来测试这项创新技术， 巴西寨卡病毒爆发的中心直接访问临床队列和患者样本， ZIKV诊断开发团队。在鉴定ZIKV反应性类肽后，我们将进行一系列分析。 免疫球蛋白对所选类肽的反应动力学的详细表征。的 这些类肽生物标志物的鉴定将导致第二代 组合文库，其中类肽反应性将进一步优化，我们将确定 ZIKV特异性类肽生物标志物与患者抗体之间的结合。本报告所述的目标 该项目将导致开发一种高度优化的，低成本的，基于类肽的高通量检测方法， 准确检测ZIKV抗体，从而诊断个体患者并测量受影响患者的发病率。 人口。这种诊断测定和疾病监测工具的开发将大大增加对疾病的诊断。 全世界公共卫生机构监测全球重要传染病的能力。