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

1449630 WiggintonEAGER：用新型小鼠模型对人诺如病毒的剂量反应消毒曲线人诺如病毒在美国引起的胃肠道疾病比任何其他病原体都多20倍，并且与这些疾病相关的年度成本估计为数十亿（美元）。尽管如此，人类诺如病毒并不包括在大多数饮用水和水再利用法规中。这在很大程度上是由于人诺如病毒不能在体外培养，即没有细胞系可用于计数样本中感染性病毒的数量。因此，科学界并不知道消灭人类诺如病毒所需的消毒处理的程度。我们目前对必要的消毒剂量和消毒动力学的理解主要依赖于替代病毒研究;尽管研究表明替代病毒对消毒剂的反应与人诺如病毒不同。饮用水和水回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合规水平。人类诺如病毒引起的死亡在发展中国家特别常见;因此，改善人类诺如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的消毒策略，最终可以预防数百万诺如病毒病例并挽救数千人的生命。PI将把这个项目纳入其正在进行的更广泛的影响活动中，包括为贫困学生开展的夏季研究活动和发展中国家的饮用水项目。在本项目中，PI建议根据感染性建立人类诺如病毒的第一条消毒曲线。密歇根大学的Christiane Wobus博士最近发现了一种被人类诺如病毒感染的免疫缺陷小鼠品系，因此代表了人类诺如病毒的第一个小动物模型。这种小鼠模型使我们能够首次开发出人类诺如病毒与消毒剂（水和废水处理中常用的化学品）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，常规监测不能基于小鼠模型，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。一旦在本项目中建立了诺如病毒的消毒曲线，我们将使用它们来校准基于定量PCR（qPCR）估计人诺如病毒灭活的方法。许多研究报告称，qPCR不能用于确定病毒感染性;然而，我们的研究推翻了这一观点，表明qPCR分析实际上可以准确测量病毒灭活，前提是在正确的理论框架内分析结果。将小鼠模型消毒数据与外推方法结合将产生简单且稳健的qPCR测定，其有效追踪人诺如病毒感染性水平，而无需细胞培养物或小鼠模型。该项目旨在提高我国水和食品供应的安全性，并减少人类诺如病毒疾病的相关经济负担。具体而言，其原因是：本项目的任务1将首次定义实现所需人类诺如病毒灭活水平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检测方法来跟踪人诺如病毒灭活。PI相信，该项目的产出将对人类诺如病毒是主要关注点的许多行业产生变革性影响，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.