RNA viruses of pandemic potential - viral infectivity

项目摘要：艾米丽·布鲁斯，博士，研究项目负责人(RPL) 核糖核酸病毒是当前和未来主要流行病的原因，几乎没有抗病毒药物，而且有可能严重 给全球医疗体系带来压力。在过去一年，严重急性呼吸系统综合症冠状病毒-2(SARS- CoV-2)是一种正链RNA病毒，是新冠肺炎的病原体，它对一种 全球范围。虽然最近一次流感大流行(2009年)的死亡率相对较低，但出现的 一种死亡率更高的新型流感病毒株的出现将使全球医疗体系处于巨大的 压力，并且仍然是一个持续的威胁。在SARS-CoV-2(以及可能任何未来的冠状病毒或 流感大流行)，对病毒诊断检测的无与伦比的需求导致全球许多 所需的试剂。RNA提取尤其是一个瓶颈，不仅是因为缺乏 需要试剂，但也由于提取过程的成本、劳动力和执行它所需的时间。 虽然最近的紧急使用授权已被美国食品和药物管理局批准用于 商业免提取诊断测试，这些是专有系统；与开放的- 需要WHO开发的来源RT-qPCR方法。 本研究的目的是探讨与SARS-CoV病毒感染性相关和/或影响的因素。 2和流感，使用临床样本和经典病毒学技术来检查是什么使这种特殊 病毒，或特定的病人，具有传染性。 先前资助的旨在开发简化的SARS-CoV-2诊断的试点项目的扩展 为了检测，我们将扩大我们的检测范围，将临床流感样本包括在内。此外，我们还将调查一个数字 包括sgRNA、负链RNA和SARS-CoV-2RNA载量在内的因素可能是 能够预测传染性病毒的存在，并可能区分那些简单的聚合酶链式反应- 来自那些真正构成病毒传播风险的人的阳性。因为这项工作将产生一组临床 对于SARS-CoV-2和具有一系列传染性的流感的分离株，我们还将调查各种 影响因素包括RNA包装、颗粒与PFU的比例、温度稳定性、颗粒形态和比 基因突变，所有这些都可能在分子水平上影响病毒的传染性。

PROJECT SUMMARY: Emily Bruce, PhD, Research Project Leader (RPL) RNA viruses are the cause of major current and future pandemics, with few antivirals and the potential to severely strain global healthcare systems. In the past year, severe acute respiratory syndrome coronavirus-2 (SARS- CoV-2), a positive strand RNA virus that is the causative agent of COVID-19, has caused acute disruption on a global scale. While the most recent influenza pandemic (2009) had a relatively low mortality rate, the emergence of a novel influenza strain with higher mortality would place the global healthcare system under immense pressure, and remains a constant threat. In the case of SARS-CoV-2 (and likely any future coronavirus or influenza pandemic), an unparalleled demand for viral diagnostic testing resulted in global shortages of many of the required reagents. RNA extraction in particular represents a choke point, not only due to shortages of the required reagents, but also due to the cost of the extraction process, the labor, and time required to perform it. While recent Emergency Use Authorizations have been approved by the US Food and Drug Administration for commercial extraction-free diagnostic tests, these are proprietary systems; protocols that work with the open- source RT-qPCR assay developed by the WHO are needed. The goal of this study is to investigate factors that correlate with and/or influence viral infectivity of SARS-CoV- 2 and influenza, using clinical samples and classical virology techniques to examine what makes the particular virus, or a particular patient, infectious. An extension of a previously funded pilot project aimed at developing a streamlined SARS-CoV-2 diagnostic test, we will expand our examination to include clinical influenza samples. Additionally, we will probe a number of factors including sgRNA, negative strand RNA and SARS-CoV-2 RNA loads that considered together may be able predict the presence of infectious virus, and potentially differentiate those individuals who are simply PCR- positive from those who truly pose a risk of viral transmission. Since this work will generate a panel of clinical isolates for both SARS-CoV-2 and influenza with a range of infectivities, we will also investigate a variety of factors including RNA packaging, particle to PFU ratio, temperature stability, particle morphology and specific genetic mutations, all of which may affect viral infectivity at the molecular level.