BRIGE: A reductionist approach to enterovirus disinfection

主要研究者：Wigginton提案编号：1228076本BRIGE提案描述了一项计划，该计划致力于对消毒过程中肠道病毒颗粒中发生的化学反应进行还原分析。该项目代表了在马里兰州大学建立一个研究小组的长期目标的第一步，该研究小组致力于采用最先进的分析技术来检测和表征病原微生物。 为了了解病毒复合体中发生的反应，必须首先了解蛋白质和基因组亚组分的反应性。虽然蛋白质和核酸单体与常见消毒剂之间的反应已得到很好的表征，但对高阶组织对单体反应性的作用知之甚少。 建议开发一种自下而上的方法来描述脊髓灰质炎病毒颗粒与游离氯的反应性。将开发定量基质辅助激光解吸电离质谱（MALDI-MS）技术，以确定氯与结构复杂性增加的生物聚合物（即，肽、RNA寡聚物、蛋白质、基因组、衣壳、感染性病毒）。在这样做时，病毒的分子结构在其消毒敏感性的作用的基本问题将得到解决。将比较三种相似脊髓灰质炎病毒株的反应性和灭活;这将阐明病毒序列的微小变化如何影响其基本反应性和消毒动力学。这项建议的扩大参与方面是与研究相结合的，它强调保留工程学科的女本科生。具体来说，本科研究项目将集中在简单的生物聚合物反应性实验。本科研究员将与马里兰州妇女工程研究员计划大学合作挑选和指导。 该提案的扩大参与方面还包括与UMD新的NSF STEP计划一起计划的活动，名为成功的工程教育和发展支持（SEEDS）。通过SEEDS，学生将接触到工程学中当前的病原体研究课题（例如，生物传感器，发展中国家的努力等）通过午餐时间的讲座和实验室参观图尔斯。建议工作的智力价值：这项研究计划将大大提高对病毒颗粒在用次氯酸消毒时发生的反应的理解。 到目前为止，科学家仍然缺乏对病毒灭活的机制描述，这项工作将作为对灭活和不灭活的修饰的整体检查。 这项工作将带来一系列科学进步，包括开发用于研究病毒蛋白质和基因组的定量MALDI-MS技术，对蛋白质和基因组组织在其对氧化剂敏感性中的作用的新视角，深入评估RNA氧化反应，和工具来预测新出现的病毒株的消毒动力学将如何不同于熟悉的病毒株的消毒动力学，拟议工作的更广泛影响：本文所述的研究工作在水卫生领域具有广泛的潜在影响。 新出现的病毒不断给环境工程师和公共卫生科学家带来新的挑战。 更好地理解突变对病毒“耐寒性”的影响，将有助于开发用于消毒工艺设计的敏感性预测工具。 此外，对病原体消毒的更好理解可能会导致有效的低成本，低能耗，消毒策略的发展。

PI: WiggintonProposal Number: 1228076This BRIGE proposal describes a plan dedicated to a reductionist analysis of the chemical reactions that take place in an enterovirus particle during disinfection. The project represents the first step of a long-term goal to develop a research team at the University of Maryland dedicated to employing state-of the art analytical techniques to detect and characterize pathogenic microorganisms. In order to understand the reactions that take place in a virus complex, one must first understand the reactivity of the protein and genome subcomponents. Although the reactions between protein and nucleic acid monomers and common disinfectants are well characterized, the role that higher order organization has on monomer reactivity is poorly understood. It is proposed that a bottom-up approach to describe a poliovirus particle's reactivity with free chlorine be developed. A quantitative Matrix Assisted Laser Desorption Ionization mass spectrometry (MALDI-MS) technique will be developed to determine the kinetics and products of reactions between chlorine and biopolymers with increasing structure complexity (i.e., peptides, RNA oligomers, proteins, genomes, capsids, infective viruses). In doing so, fundamental questions about the role of virus molecular structure in its disinfection susceptibility will be addressed. The reactivity and inactivation of three similar poliovirus strains will be compared; this will elucidate how slight changes in a virus sequence influence its fundamental reactivity and disinfection kinetics. The broadening participation aspect of this proposal is integrated with the research-it emphasizes retaining female undergraduate students in engineering disciplines. Specifically, an undergraduate research project will focus on the simple biopolymer reactivity experiments. The undergraduate researcher will be selected and mentored in collaboration with the University of Maryland Women in Engineering Fellows Program. Also included in the broadening participation aspect of this proposal are planned activities with the new NSF STEP program at UMD titled Successful Engineering Education and Development Support (SEEDS). With SEEDS, students will be exposed to current pathogen research topics in engineering (e.g., biosensors, efforts in the developing world, etc.) through lunchtime lectures and lab tours. Intellectual Merit of Proposed Work: This research plan will substantially improve the understanding of the reactions that take place in a virus particle as it is disinfected with hypochlorous acid. To date, scientists continue to lack a mechanistic description of virus inactivation and this work will serve as a holistic examination of the modifications that do and do not contribute to inactivation. The work will result in a number of scientific advancements including the development of a quantitative MALDI-MS technique for studying virus proteins and genomes, a new perspective on the role of protein and genome organization in their susceptibility to oxidants, an in-depth evaluation of RNA oxidation reactions, and tools to predict how the disinfection kinetics of emerging virus strains will differ from the disinfection kinetics of familiar virus strains based on nucleic acid and amino acid mutations.Broader Impacts of Proposed Work: The research effort described herein has wide reaching potential impacts in the field of water sanitation. Emerging viruses constantly create new challenges for environmental engineers and public health scientists. A better comprehension of the influence that mutations have in virus "hardiness" will allow for the development of susceptibility prediction tools for use in disinfection process design. Furthermore, an improved understanding on pathogen disinfection could lead to the development of effective low-cost, low-energy, disinfecting strategies.