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

项目总结/摘要 新出现的传染病对人类健康构成重大风险。在新出现的病原体中， 出血热病毒（HFV）在潜在的发病率方面构成最大的风险， mortality.这些病毒在偏远地区流行，产生大规模爆发， 死亡率很高，很有可能成为流行病。HFV最大的家族之一是 沙粒病毒，其中包括6种DHHS选择因子。其中最著名的是拉沙病毒， 近年来导致越来越多的感染，包括2018年在尼日利亚爆发， 造成了100多人死亡2017年，世界卫生组织将拉沙病毒列为优先事项。 病原体的研发工作，因为它有可能产生公共卫生紧急情况。其他出血性 包括Lujo、Chapare、Guanarito、朱宁和Machupo在内的发热沙粒病毒代表了不断增加的 由于其分布增加和最近发生的人与人之间的冲突， 传输目前对拉沙病毒和其他沙粒病毒的诊断能力极其有限。 有限，特别是在护理点（POC）诊断测定的情况下。在这里，我们建议制定一个 具有诊断拉沙病毒所需的灵敏度和覆盖范围的POC诊断测定 目前在西非存在感染。此外，我们将产生单链抗体噬菌体 展示库，广泛反应的高亲和力诊断抗体对，用于5 A类优先病原体的其他沙粒病毒。我们以前使用过这些程序 开发一种针对埃博拉病毒的POC诊断检测方法，其灵敏度高于目前的黄金检测方法。 标准品，PCR。这些研究的成功完成将导致第一次POC诊断检测 能够检测所有具有临床重要性的拉沙菌株， 所有A类沙粒病毒的抗体和检测。此外，广泛反应性单链抗体 我们产生的文库可以用来快速产生针对新型沙粒病毒的诊断抗体 动物传染病的爆发。因此，这项工作将大大改善我们对几乎所有国家的准备工作。 任何未来可能爆发大规模流行的沙粒病毒。

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.