Development of point-of-care testing for Lassa and other hemorrhagic fever arenaviruses

项目摘要/摘要 新出现的传染病对人类健康构成重大威胁。在新出现的病原体中， 出血热病毒(HFV)在发病和死亡的可能性方面构成最大的风险 死亡率。这些病毒在偏远地区是地方性的，会产生大规模的疫情，表现出很高的 死亡率，并有很高的潜在成为大流行。出血热病毒最大的家族之一是 ArenaVirus，其中包括6种DHHS精选药物。其中最广为人知的拉萨病毒 近年来导致感染人数不断增加，包括2018年在尼日利亚爆发的导致 超过100人死亡。2017年，世界卫生组织将拉萨病毒列为优先事项 由于其可能引发公共卫生紧急情况，因此可能会对研发工作造成不利影响。其他出血性疾病 包括Lujo、Chapare、Guanarito、Junin和Machupo在内的发热病毒代表着越来越多的 根据它们日益增加的分布和最近人与人之间的传播情况而引起的关切 变速箱。目前对拉萨病毒和其他阿拉伯病毒的诊断能力非常强 有限，特别是在护理点(POC)诊断分析的情况下。在这里，我们建议开发一种 POC诊断试验，具有诊断拉萨病毒所需的灵敏度和覆盖面 目前在西非存在感染。此外，我们还将产生单链抗体噬菌体 展示文库、广谱反应性高亲和力诊断抗体对、5个原型化验芯片 属于A类优先病原体的其他阿拉伯病毒。我们以前曾使用过这些程序 开发一种敏感性高于目前金标的埃博拉病毒POC诊断方法 标准，聚合酶链式反应。这些研究的成功完成将导致第一个POC诊断化验 它能够检测所有临床上重要的拉沙菌菌株和一组高亲和力的诊断 所有A类禽流感病毒的ABS和化验。此外，广谱反应性单链抗体 我们生产的文库可以用来快速产生针对新型阿拉伯病毒的诊断抗体 以人畜共患病疫情的形式出现。因此，这项工作将大大改善我们对 未来任何具有大流行潜力的主要禽流感疫情。

Project Summary/Abstract Emerging infectious diseases pose a significant risk to human health. Among emerging pathogens, hemorrhagic fever viruses (HFVs) pose the greatest risk in terms of the potential for morbidity and mortality. These viruses are endemic in remote regions, produce large outbreaks that display high mortality rates, and have a high potential become pandemics. One of the largest families of HFVs is the Arenaviruses, which includes 6 DHHS select agents. The most well-known of these, Lassa virus, has caused an increasing number of infections in recent years, including a 2018 outbreak in Nigeria that led to over 100 deaths. In 2017, the World health Organization designated Lassa virus as a priority pathogen for R&D efforts due to its potential to generate a public health emergency. Other hemorrhagic fever arenaviruses, including Lujo, Chapare, Guanarito, Junin, and Machupo represent an increasing source of concern based on their increased distribution and recent instances of human-to-human transmission. Current diagnostic capabilities for Lassa virus and other arenaviruses are extremely limited, especially in the case of point-of-care (POC) diagnostic assays. Here, we propose to develop a POC diagnostic assay with the sensitivity and breadth of coverage required to diagnose Lassa virus infections currently present in West Africa. In addition, we will generate single chain antibody phage display libraries, broadly reactive high affinity diagnostic antibody pairs, a prototype assay chips for the 5 other arenaviruses that are Category A Priority Pathogens. We have previously used these procedures to develop a POC diagnostic assay for Ebola virus that displays a sensitivity better than the current gold standard, PCR. The successful completion of these studies will result in the first POC diagnostic assay that is capable of detecting all Lassa strains of clinical importance and a panel of high affinity diagnostic Abs and assays for all Category A Arenaviruses. In addition, the broadly reactive single chain antibody libraries we produce could be used to rapidly generate diagnostic antibodies against novel arenaviruses that arise as zoonotic outbreaks. This work will thus significantly improve our preparedness for almost any future major Arenavirus outbreak with pandemic potential.