BRIGE: A reductionist approach to enterovirus disinfection

提案编号：1228076本bridge提案描述了一个致力于对肠道病毒颗粒在消毒过程中发生的化学反应进行还原分析的计划。该项目是马里兰大学长期目标的第一步，该目标是建立一个研究团队，致力于采用最先进的分析技术来检测和表征病原微生物。为了理解病毒复合体中发生的反应，我们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普通消毒剂之间的反应已经被很好地表征，但高阶组织对单体反应性的作用却知之甚少。提出了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。将开发一种定量基质辅助激光解吸电离质谱（MALDI-MS）技术，以确定氯与结构日益复杂的生物聚合物（即肽、RNA寡聚物、蛋白质、基因组、衣壳、感染性病毒）之间反应的动力学和产物。在此过程中，有关病毒分子结构在其消毒敏感性中的作用的基本问题将得到解决。将比较三种类似脊髓灰质炎病毒株的反应性和灭活性；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。该提案的扩大参与方面与研究相结合，它强调在工程学科中保留女性本科生。具体来说，本科生的研究项目将侧重于简单的生物聚合物反应性实验。本科研究人员将与马里兰大学女性工程研究员项目合作选择和指导。该提案还包括扩大参与方面的内容，包括与UMD新的NSF STEP项目一起计划的活动，名为“成功的工程教育和发展支持”（SEEDS）。有了SEEDS，学生将通过午餐讲座和实验室参观，接触到当前工程中的病原体研究主题（例如，生物传感器，在发展中国家的努力等）。建议工作的智力价值：本研究计划将大大提高对用次氯酸消毒病毒颗粒时发生的反应的理解。迄今为止，科学家们仍然缺乏对病毒灭活的机制描述，这项工作将作为对那些有助于和不有助于灭活的修饰的全面检查。这项工作将带来许多科学进步，包括开发用于研究病毒蛋白质和基因组的定量MALDI-MS技术，从新的角度研究蛋白质和基因组组织在它们对氧化剂的易感性中的作用，深入评估RNA氧化反应，以及预测新出现病毒株的消毒动力学与基于核酸和氨基酸突变的熟悉病毒株的消毒动力学有何不同的工具。拟议工作的更广泛影响：本文所描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“抗寒性”的影响将有助于开发用于消毒过程设计的易感性预测工具。此外，提高对病原体消毒的认识可能会导致开发有效的低成本，低能耗的消毒策略。

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.