Vesicle-Cloaked Virus Clusters as Emerging Pathogens: Will They Challenge Current Disinfection Paradigm?

项目总结/摘要 众所周知，轮状病毒和诺如病毒会导致人类严重的胃肠炎，并极大地影响公共卫生 性问题这两种病毒都具有环境持久性，它们可以通过直接接触传播。 受感染的人或受污染的表面，以及受污染的水和食物。消毒可以消除这些 持久性病毒和预防传染病爆发。几乎所有以前的研究都认为轮状病毒和 在生物医学研究中，诺如病毒作为单独的、游离的病毒颗粒，直到最近的一项发现表明， 轮状病毒和诺如病毒可以在体外作为病毒簇在细胞外囊泡内传播， 膜结构，即，囊泡包被的病毒簇（简单来说是病毒囊泡）。囊泡包被轮状病毒 诺如病毒簇是重要的新兴病原体，因为它们在环境中稳定， 存在，并且比它们的游离病毒颗粒对应物更具感染性;然而，它们被电流灭活， 消毒策略尚不清楚。此外，消毒机制在很大程度上还不清楚，这使我们无法 从有效的，强大的，广谱的，低成本的消毒过程的合理设计。项目的目标是 评价灭活囊泡包被轮状病毒的常规消毒策略的性能， 诺如病毒簇，阐明病毒囊泡灭活机制，并开发新的消毒剂 用于有效的病毒囊泡灭活的方法。我们的核心假设是囊泡病毒聚集在 与游离病毒的对应物相比， 病毒囊泡中的多种生物成分更有效。具体目标1将评价以下物质的灭活 常规消毒剂，即，漂白剂，紫外光（单色UV 254 nm）、表面活性剂（二癸基二甲基氯化铵）、乙醇和它们的组合。的剂量 将确定消毒剂及其消毒性能以产生剂量-反应关系， 这将与自由病毒的情况进行比较。具体目标2将阐明失活的机制 泡状病毒群，以了解决定 消毒性能病毒囊泡生物组分的损伤及其对病毒囊泡寿命的影响 将评价消毒后宿主细胞中的生命周期。具体目标3将评估新兴的消毒剂， 也就是说，多色紫外线照射和活化过硫酸盐，用于灭活囊泡包裹的病毒簇。两 消毒性能和机制将被探索。我们的研究将阐明优化的必要性 当前的消毒范式并开发针对轮状病毒和诺如病毒的新消毒策略，并且它已经 更广泛的健康影响，因为研究成果可以转化为灭活其他相关病毒 (e.g.,脊髓灰质炎病毒、柯萨奇病毒、鼻病毒、甲型和戊型肝炎病毒），这些病毒具有环境持久性， 也能形成囊泡状病毒簇。

Project Summary/Abstract Rotavirus and norovirus are well-known to lead to serious gastroenteritis in humans and great public health concerns. Both viruses are environmentally persistent, and they can be transmitted via direct contact with an infected person or contaminated surfaces, as well as polluted water and food. Disinfection can inactivate these persistent viruses and prevent infectious disease outbreaks. Almost all previous studies consider rotavirus and norovirus as individual, free viral particles in biomedical research, until a recent finding demonstrated that rotavirus and norovirus could transmit in vitro as viral clusters inside extracellular vesicles that have a lipid-rich membrane structure, i.e., vesicle-cloaked virus clusters (viral vesicles for simplicity). Vesicle-cloaked rotavirus and norovirus clusters are important emerging pathogens, because they are environmentally stable, widely present, and more infectious than their free viral particle counterpart; however, their inactivation by current disinfection strategies is unknown. Moreover, disinfection mechanisms are largely unclear, which prevents us from rational design of effective, robust, broad-spectrum, and low-cost disinfection processes. This project seeks to evaluate the performance of conventional disinfection strategies for inactivating vesicle-cloaked rotavirus and norovirus clusters, elucidate the mechanisms of viral vesicle inactivation, and develop new disinfection processes for effective viral vesicle inactivation. Our central hypothesis is that the vesicle-cloaked virus clusters are more persistent in disinfection compared to their counterpart of free viruses, and disinfection that damages multiple biological components in viral vesicles is more effective. Specific Aim 1 will evaluate the inactivation of vesicle-cloaked virus clusters by conventional disinfectants, i.e., bleach, ultraviolet light (monochromatic UV 254 nm), a surfactant (didecyldimethylammonium chloride), ethanol, and their combination. The dosage of disinfectants and their performance in disinfection will be determined to generate a dose-response relationship, which will be compared with that for free viruses. Specific Aim 2 will elucidate the mechanisms of inactivating vesicle-cloaked virus clusters by conventional disinfectants to understand the key factor that determines disinfection performance. The damage of biological components of viral vesicles and the impact on viral vesicles’ lifecycle in the host cells after disinfection will be evaluated. Specific Aim 3 will evaluate emerging disinfectants, i.e., polychromatic UV irradiation and activated persulfate, for inactivating vesicle-cloaked virus clusters. Both disinfection performance and mechanisms will be explored. Our study will shed light on the need for optimizing current disinfection paradigm and developing new disinfection strategies for rotavirus and norovirus, and it has a broader health impact because research outcomes can be translated into inactivating other related viruses (e.g., poliovirus, coxsackievirus, rhinovirus, hepatitis A and E viruses), which are environmentally persistent and could also form vesicle-cloaked virus clusters.