UNS: Mechanisms of UV inactivation of viruses for tailored disinfection applications

[12616 . rodriguez]这项建议的目标是获得紫外线对病毒作用的基本见解。该研究将研究分子水平上病毒对紫外线照射的反应，以改进当前的紫外线消毒和设计未来的紫外线技术。该研究旨在回答有关紫外线灭活病毒的一些基本问题，这些问题可以为工程实践、公共卫生和随后的监管政策提供信息，从而具有广泛的影响。拟议的活动将促进若干关键领域的知识。新的基础知识将发展到病毒在消毒实践中对紫外线照射的分子水平反应。具体来说，除了腺病毒外，还将对另一种双链DNA病毒（JC多瘤病毒）进行新的研究，以测试腺病毒对低压和中压紫外线的反应结果的普遍性。这项研究将超越对实验室腺病毒株的传统评估，并分离和测试环境菌株，以确定在实验室细胞培养系中反复暴露和繁殖时，是否存在与适应反应（如DNA修复）有关的紫外线反应的任何差异。分子微生物学方法的许多新领域，其中一些是我们开发的，将被开发，包括由于特定紫外线波长的早期感染抑制，跨病变合成的DNA修复，使用电泳和质谱技术的蛋白质损伤评估，以及使用我们最近发表的新的远程PCR技术的DNA损伤。这些基本发现将用于开发一种基于定制波长紫外led的节能照射系统，以优化消毒，比传统紫外线源成本更低。本文提出的工作将：(1)评估两种双链DNA病毒的失活动力学和波长敏感性，(2)将腺病毒的研究扩展到从环境中分离的腺病毒，(3)评估DNA修复在细胞培养感染性结果中的基本作用，(4)结合新开发的基于分子生物学的检测方法来评估不同波长紫外线照射对DNA和蛋白质的损伤，以及，(5)了解纳米特异性紫外线发光二极管（led）的发展对使用定制波长方法进行消毒的影响。监管机构和公共卫生官员需要信息来帮助公共部门做出合理的决策。小型系统的病毒消毒问题目前存在争议，因为地下水规则明确禁止使用紫外线进行任何病毒灭活，因为腺病毒灭活所需的高剂量目前无法在紫外线反应器中进行现场验证。因此，如果紫外线消毒将对地下水中的病毒消毒发挥任何作用，这项研究的时机至关重要。因此，想要利用紫外线保护公众健康的小型系统可能会遭受不必要的损失。

1512616RodriguezThe goal of this proposal is to gain fundamental insights into the action of ultraviolet light on viruses. The study will examine the molecular level viral responses to ultraviolet irradiation to improve current ultraviolet light disinfection and design of future ultraviolet light technologies. The research aims to answer a number of fundamental questions around virus inactivation with ultraviolet light that can inform engineering practice, public health, and subsequent regulatory policy and thus has broad reaching impacts.The proposed activity will advance knowledge in a number of key areas. New fundamental knowledge will be developed into the molecular-level response of viruses to ultraviolet light irradiation during disinfection practice. Specifically, in addition to adenoviruses, new studies will be performed on another double stranded DNA virus - JC polyomavirus - to test the generalizability of the findings with adenoviruses in response to low-pressure and medium-pressure ultraviolet light. This research will move beyond the traditional evaluations of laboratory strains of adenoviruses and isolate and test environmental strains to determine if there are any differences in the ultraviolet light -response that may be related to acclimated responses (such as DNA repair) from repeated exposure to and propagation in laboratory cell culture lines. A number of new areas in molecular microbiology methods, some of which we have developed, will be exploited including early stage infection inhibition due to specific ultraviolet light wavelengths, DNA repair from trans-lesion synthesis, protein damage assessment using electrophoresis and mass spectrometry techniques, and DNA damage using a new long range PCR technique we recently published. These fundamental findings will be used to develop a tailored wavelength ultraviolet-LED based energy efficient irradiation system to optimize disinfection at lower cost than conventional ultraviolet sources. The work proposed herein will: (1) evaluate inactivation kinetics and wavelength sensitivity of two double stranded DNA viruses, (2) extend the adenovirus studies to adenoviruses isolated from the environment, (3) evaluate the fundamental role of DNA repair in the cell culture infectivity outcomes, (4) incorporate newly developed molecular biology-based assays to assess DNA damage and protein damage from ultraviolet light irradiation at varying wavelengths, and, (5) understand the implications for disinfection using tailored wavelength approaches made possible by the development of nanometer-specific ultraviolet light emitting diodes (LEDs). Regulators and public health officials need information to help make sound decisions for the public sector. The issue of virus disinfection for small systems is currently contentious as the Groundwater Rule specifically forbids the use of UV for any virus inactivation credit due to the high doses required for adenovirus inactivation that cannot currently be field-validated in UV reactors. Therefore, the timing of this research is critical if UV disinfection is going to play any role for virus disinfection in groundwater. Thus, small-systems wanting to use UV for protection of public health, may needlessly suffer.