The development of a rapid diagnostic test for Ebola virus.

2014- 2015年西非埃博拉疫情暴露了世界对传染病的脆弱性，今天埃博拉仍然是世卫组织具有高流行潜力和仍然需要研发的重点病原体清单的首位。具体而言，仍然迫切需要快速诊断测试，因为目前的工具缺乏检测泛埃博拉物种的敏感性和特异性，以及支持分散检测的数据连通性。该博士项目的目的是建立新一代快速、低成本和易于操作的诊断测试，用于埃博拉病毒病（EVD）的早期诊断，能够在分散的资源贫乏环境中检测不同的病毒种类。该项目将以LCN的McKendry小组和美国陆军传染病医学研究所（USAMRIID）的Dye小组之间正式建立的合作为基础。该项目建立在Dye博士在《细胞》杂志上发表的一项重大突破的基础上，该突破筛选了来自人类幸存者的349种单克隆抗体，并确定了可特异性识别泛埃博拉物种的主要候选抗体，目标是GP融合环中的启动子间表位，这是病毒膜融合机制的关键和保守元件。在这个项目中，Dye博士将分享他在内部生产的针对埃博拉病毒和马尔堡病毒的独特单克隆抗体，该抗体具有高特异性和敏感性，可识别特定的病毒蛋白。这些捕获抗体的特征及其与纳米颗粒的结合将使用ELISA和生物层干涉法进行。然后，将鉴定出的抗体用于开发多目标分析物横向流动试纸条，用于检测和区分埃博拉病毒毒株，包括苏丹病毒、扎伊尔病毒和本迪布焦病毒。该测试条将由多个测试线和修改的纳米颗粒组成，用于量热结果读出。然后，将使用重组病毒蛋白对分析进行校准，并确定检测限。最后，新出现的传染病与恐怖主义等威胁并列，在内阁办公室风险登记册中名列前茅。此外，该项目提案与UKRI、GCRF、NIHR、联合国和世卫组织在传染病快速诊断检测和应急准备方面的优先事项非常一致。因此，成功开发这种针对急性埃博拉感染的快速检测方法可以为全世界的患者和人群带来重大的人类和经济利益

The 2014-15 Ebola in West Africa outbreak exposed the world's vulnerabilities to infectious diseases and today Ebola remains at the top of the WHO list for priority pathogens with high epidemic potential and remaining need for R&D. Specifically, there is still an urgent need for rapid diagnostic tests as current tools lack the sensitivity, specificity to detect pan-Ebola species and data connectivity to support decentralised testing. The aim of this PhD project is to build a new generation of rapid, low cost and easy to operate diagnostic test for early diagnosis of Ebola virus disease (EVD), with the ability to detect different viral species, in decentralised resource-poor settings. This project will be based on a formally established collaboration between McKendry's group at LCN and Dye's group at the U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID). This project builds on a major breakthrough by Dr Dye published in the journal "Cell" which screened 349 monoclonal antibodies from human survivors and identified lead candidates which can specifically identify pan-Ebola species, targeting the inter-promoter epitope in the GP fusion loop, a critical and conserved element of the viral membrane fusion machinery. For this project Dr Dye will share his in-house production of unique monoclonal antibodies against Ebolavirus and Marburgvirus with high specificity and sensitivity to identify specific viral proteins. Characterisation of these Capture antibodies and their conjugation to nanoparticles will be performed using ELISA and Biolayer-Interferometry. Then, the characterized antibodies will be used for the development of multiple target analyte lateral flow test strip for the detection and differentiation of Ebola virus strains, including Sudan virus, Zaire virus and Bundibgyo virus. This test strip will be comprised of multiple test lines and modified nanoparticles for calorimetric result readout. Then, the assay will be calibrated and the limit of detection will be determined using recombinant viral proteins. Finally, emerging infections rank at the top of over Cabinet Office Risk Register, alongside threats such as terrorism. Moreover, this project proposal strongly aligns to UKRI, GCRF, NIHR, UN and WHO priorities for rapid diagnostic tests for infectious diseases and emergency preparedness. Hence, successful development of this rapid tests for acute Ebola infections could contribute to major human and economic benefits to patients and populations worldwide