EAGER: Dose-Response Disinfection Curves for Human Norovirus with Novel Mouse Model

1449630WiggintonEAGER：人类诺沃克病毒与新型小鼠模型的剂量-反应消毒曲线人类诺沃克病毒引起的胃肠道疾病大约是美国任何其他病原体的20倍，与这些疾病相关的年成本估计高达数十亿美元(美元)。尽管如此，人类诺沃克病毒并不包括在大多数饮用水和水重复使用法规中。这在很大程度上是因为人类诺沃克病毒不能在体外培养，也就是说，没有细胞株可以用来统计样本中感染病毒的数量。因此，科学界不知道灭活人类诺沃克病毒所需的消毒治疗的程度。我们目前对必要的消毒量和消毒动力学的了解主要依赖于代理病毒的研究；尽管研究表明，代理病毒对消毒剂的反应与人类诺沃克病毒不同。饮用水和水再利用领域迫切需要准确的人类诺如病毒消毒数据，以便建立水处理指南和遵从性水平。人类诺沃克病毒造成的死亡在发展中国家尤其常见；因此，改进人类诺沃克病毒消毒将对贫困人口产生积极影响。最终，这项研究将导致改进消毒策略，最终可能防止数百万诺沃克病毒病例，并拯救数千人的生命。私营部门将把这一项目纳入其正在进行的更广泛的影响活动中，包括针对贫困学生的暑期研究活动和发展中国家的饮用水项目。在这个项目中，PI建议根据传染性建立第一个人诺沃克病毒的消毒曲线。密歇根大学的克里斯蒂安·沃布斯博士最近发现了一种免疫缺陷小鼠品系，该品系感染了人类诺沃克病毒，从而代表了人类诺沃克病毒的第一个小动物模型。这个小鼠模型使我们第一次能够用消毒剂(水和废水处理中常用的化学物质)绘制人类诺沃克病毒的剂量反应曲线。这些消毒剂量-反应曲线是至关重要的；然而，常规监测不能基于老鼠模型，而必须依赖于更大规模的科学家和从业者群体可用的更便宜的技术。一旦在这个项目中建立了诺沃克病毒的消毒曲线，我们将使用它们来校准一种基于定量聚合酶链式反应(QPCR)估计人类诺沃克病毒灭活的方法。数十项研究报告称，qPCR不能用于确定病毒的传染性；然而，我们的研究推翻了这一概念，表明如果在正确的理论框架内分析结果，qPCR分析实际上可以准确地测量病毒灭活。将小鼠模型消毒数据与外推方法相结合，将产生简单而可靠的qPCR分析，有效地跟踪人类诺沃克病毒的传染性水平，而不需要细胞培养或小鼠模型。该项目旨在提高我国水和食品供应的安全性，并减少与人类诺沃克病毒疾病相关的经济负担。特别是，本项目任务1的结果将首次确定实现所需水平的人类诺沃克病毒灭活所需的特定紫外线剂量。任务2将介绍一种简单且廉价的qPCR方法来跟踪人类诺沃克病毒的灭活。私人投资机构相信，该项目的成果将对一些主要关注人类诺沃克病毒的行业产生革命性影响，包括饮用水、废水、食品和医院。

1449630WiggintonEAGER: Dose-response disinfection curves for human norovirus with novel mouse modelHuman norovirus causes approximately 20 times more gastrointestinal illnesses than any other pathogenic agent in the U.S., and the annual cost associated with these illnesses is estimated to be in the billions (US $). Despite this, human norovirus is not included in most drinking water and water reuse regulations. This is largely due to the fact that Human Norovirus cannot be cultured in vitro, that is, no cell line is available to count the number of infective viruses in samples. As a result, the scientific community does not know the extent of disinfecting treatments necessary to inactivate human norovirus. Our current understanding of necessary disinfection doses and disinfection kinetics relies primarily on surrogate virus studies; this is despite the fact that research has indicated that the surrogate viruses respond differently to disinfectants than human norovirus. The drinking water and water reuse fields are in desperate need of accurate human norovirus disinfection data so that water treatment guidelines and compliance levels can be established. Deaths caused by human norovirus are especially common in the developing world; improved human norovirus disinfection would therefore have positive impacts on impoverished populations. Ultimately, the research will lead to improved disinfection strategies, which could ultimately prevent millions of norovirus cases and save thousands of lives. The PIs will incorporate this project into their ongoing broader impact activities, including summer research activities for underprivileged students and drinking water projects in developing countries. In this project, the PIs propose to establish the first disinfection curves of human norovirus based on infectivity. Dr. Christiane Wobus of the University of Michigan recently discovered an immune deficient mouse strain that is infected by human norovirus, and thus represents the first small animal model for human norovirus. This mouse model allows us for the first time to develop dose-response curves for the human norovirus with disinfectants (chemicals commonly used in water and wastewater treatment). These disinfection dose-response curves are critical; however, routine monitoring cannot be based on the mouse model, and must instead rely on cheaper techniques available to a much larger group of scientists and practitioners. Once disinfection curves of norovirus have been established in this project, we will use them to calibrate a method that estimates human norovirus inactivation based on quantitative PCR (qPCR). Scores of studies have reported that qPCR cannot be used to determine virus infectivity; however our research has overturned this notion by showing that qPCR analysis can, in fact, accurately measure virus inactivation, provided that the results are analyzed within the correct theoretical framework. Coupling the mouse model disinfection data with the extrapolation method will lead to a simple and robust qPCR assay that effectively tracks human norovirus infectivity levels without the need for cell cultures or the mouse models. This project aims to increase the safety of our nation's water and food supplies and reduce the associated economic burden of human norovirus illnesses. In particular, results from: Task 1 of this project will, for the first time, define specific UV doses necessary to achieve desired levels of human norovirus inactivation. Task 2 will introduce a simple and inexpensive qPCR assay to track human norovirus inactivation. The PIs are confident that the outputs from this project will be transformative for a number of industries for which human norovirus is a major concern including drinking water, wastewater, food, and hospital.