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

项目摘要/摘要 众所周知，轮状病毒和诺沃克病毒会导致人类严重的胃肠炎和极大的公共健康 担忧。这两种病毒在环境中都是持久性的，它们可以通过与 感染者或受污染的表面，以及受污染的水和食物。消毒可以使这些细胞失活 持久的病毒，防止传染病暴发。几乎所有以前的研究都认为轮状病毒和 诺沃克病毒作为个体，在生物医学研究中作为游离的病毒颗粒，直到最近的一项发现证明 轮状病毒和诺如病毒可以在体外以病毒簇的形式在富含脂质的细胞外小泡内传播。 膜结构，即囊泡覆盖的病毒簇(简单地说，病毒囊泡)。水泡衣状轮状病毒 而诺沃克病毒簇是重要的新兴病原体，因为它们在环境中稳定，广泛 存在，并且比它们的游离病毒颗粒更具传染性；然而，它们通过电流灭活 消毒策略尚不清楚。此外，消毒机制在很大程度上不清楚，这阻止了我们 从合理设计有效、坚固、广谱和低成本的消毒工艺。这个项目寻求 评价常规消毒策略对水泡包裹轮状病毒灭活效果和 诺如病毒簇，阐明病毒囊泡灭活机制，开发新的消毒 有效灭活病毒泡的方法。我们的中心假设是囊泡覆盖的病毒群 与其对应的游离病毒相比，在消毒方面更持久，而且消毒会损害 病毒囊泡中的多种生物成分更有效。特定目标1将评估失活 用常规消毒剂，即漂白剂、紫外线(单色紫外线254)覆盖水泡的病毒团 Nm)、表面活性剂(二十二烷基二甲基氯化铵)、乙醇及其组合。安慰剂用量 消毒剂及其消毒性能将被确定为产生剂量-反应关系， 这将与免费病毒的情况进行比较。具体目标2将阐明灭活的机制。 通过常规消毒剂了解水泡衣壳病毒聚集性的关键因素 消毒性能。病毒囊泡生物成分的损伤及其对病毒囊泡的影响 将对消毒后宿主细胞的生命周期进行评估。特殊目标3将评估新出现的消毒剂， 即多色紫外线照射和活化过硫酸盐，用于灭活水泡覆盖的病毒簇。两者都有 将探索消毒性能和机制。我们的研究将阐明优化的必要性 目前针对轮状病毒和诺沃克病毒的消毒范例和开发新的消毒策略，并已 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 (例如，脊髓灰质炎病毒、柯萨奇病毒、鼻病毒、甲型肝炎和戊型肝炎病毒)，这些病毒在环境上是持久的和 也可以形成囊泡覆盖的病毒簇。

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.