Point-of-care antigen detection assay for early diagnosis of Ebola virus disease (EVD)

项目摘要 埃博拉病毒属引起一种称为埃博拉病毒病（EVD）的严重出血热。EVD是 这是一个严重的公共卫生问题，既是一种新出现的传染病，也是一种潜在的 生物威胁2013-2016年西非埃博拉大流行引起了全球的关注， 与控制埃博拉病毒有关的挑战。民主党正在爆发的疫情 刚果共和国最近达到1000多例，没有减缓的迹象， 证实需要改进埃博拉医疗对策。为了更好地控制埃博拉 疫情爆发时，医护人员必须能够诊断和隔离感染者， 患者及时就医。因此，世界卫生组织和 其他医疗机构呼吁开发埃博拉快速诊断测试（RDTs）。 拟议的共同努力，为发展一个即时的抗原检测测定，为早期 EVD的诊断，源于学术界之间成功合作的历史， 里诺（UNR）内华达州大学的大卫AuCoin博士的实验室和商业 InBios International，Inc.这两个组织在RDT方面拥有宝贵的专业知识 开发和导航FDA批准程序。这一努力得到了以下方面的进一步支持： 美国陆军传染病医学研究所的合作者的帮助下， 疾病（USAMRIID）谁将执行所有高遏制水平（BSL 4）的研究。 对这一提议的初步研究已经产生了一个埃博拉RDT原型， 显著提高了对扎伊尔埃博拉病毒敏感性。我们的目标是进一步提高检测 灵敏度和扩展检测所有EVD引起的埃博拉病毒属。综合办法 有明确的战略将用于支持泛埃博拉病毒的成功发展， RDT具有高灵敏度。关键方面和里程碑包括：i）高通量单克隆抗体 使用多种免疫策略产生抗体（mAb），ii）mAb表征和 基于RDT的mAb筛选，iii）商业优化和开发多种测定 原型，iv）测定分析灵敏度和特异性，以及v）使用 感染埃博拉病毒的非人类灵长类动物的样本

Project Summary Ebolavirus spp. cause a severe hemorrhagic fever known as Ebola virus disease (EVD). EVD is a serious public health concern, both as an emerging infectious disease and a potential biothreat. The 2013-2016 Ebola pandemic in West Africa brought global attention to the challenges associated with controlling an Ebola. The ongoing outbreak in the Democratic Republic of Congo, which recently reached over 1000 cases with no signs of slowing, has only confirmed the need for improved Ebola medical countermeasures. To better control Ebola outbreaks, it is imperative for healthcare workers to be able to diagnosis and isolate infected patients at the point-of-care in a timely manner. As such, the World Health Organization and other healthcare agencies have called for development of Ebola rapid diagnostic tests (RDTs). The proposed joint effort, for development a point-of-care antigen detection assay for early diagnosis of EVD, stems from a history of successful collaborations between the academic laboratory of Dr. David AuCoin at the University of Nevada, Reno (UNR) and commercial partner, InBios International, Inc. The two organizations have valuable expertise in RDT development and navigating the FDA approval process. The effort is further bolstered by assistance from collaborators at the United States Army Medical Research Institute of Infectious Diseases (USAMRIID) who will perform all high containment level (BSL4) studies. Preliminary studies for this proposal have resulted in an Ebola RDT prototype that shows a substantial improvement in sensitivity for Zaire ebolavirus. Our goal is to further improve assay sensitivity and expand detection to all EVD causing Ebolavirus spp. A comprehensive approach with clearly defined strategies will be used to support successful development of a pan-Ebola RDT with high sensitivity. Key aspects and milestones include: i) high throughput monoclonal antibody (mAb) production using multiple immunization strategies, ii) mAb characterization and RDT-based mAb screening, iii) commercial optimization and development of multiple assay prototypes, iv) determination of analytical sensitivity and specificity and v) assay validation with samples from Ebola-infected non-human primates.