紫蓝光(405 nm)灭活SARS-CoV-2机制分析

IF 3.9 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Davide Amodeo, Serena Marchi, Lia Fiaschi, Luisa Raucci, Camilla Biba, Valentina Salvestroni, Claudia Maria Trombetta, Ilaria Manini, Maurizio Zazzi, Emanuele Montomoli, Ilaria Vicenti, Gabriele Cevenini, Gabriele Messina
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引用次数: 0

摘要

本研究评估了紫蓝光(VBL)对SARS-CoV-2细胞活力和复制、3种结构蛋白(S、E和N)和1种非结构蛋白(NSP13)羰基化的影响,以及对RNA的直接损伤。将该病毒与甲型和乙型流感病毒一起暴露于增加剂量的VBL,以比较它们的易感性。在最高剂量(21.6 J/cm2)下,SARS-CoV-2对VBL的易感性明显高于流感病毒,病毒滴度降低2.33 log10。新一代测序和实时PCR定量显示,暴露于VBL后,病毒RNA未显示出显著变化,表明失活过程不涉及直接核酸损伤。为了排除培养悬浮液在灭活过程中的作用,使用不同稀释度的Dulbecco改良Eagle培养基(DMEM)在磷酸盐缓冲盐水(PBS)中进行病毒活力实验。结果表明,悬浮培养基在病毒灭活中起次要作用,因为活力不随DMEM稀释度的增加而增加。随后用三种不同浓度的抗氧化剂(NAC、AsA和SOD)对病毒进行灭活试验,其效果从99.99%到85.43%不等(SOD为0.003 mM)。当病毒悬浮在DMEM而不是PBS中时,S和E蛋白的羰基化更为明显,尽管试验表明,病毒膜的内在特性是考虑其对VBL易感的关键因素。基于灯光的消毒方法通常与其他清洁方法结合使用,因为它们具有非侵入性、多功能性和环境效益。VBL是一种有效的方法,因为它可以诱导活性氧的产生,从而降低微生物的生存能力。在这项研究中,脂质过氧化被认为是影响病毒包膜结构完整性和功能的重要因素,降低了其与宿主细胞相互作用的能力,从而降低了其感染能力。SARS-CoV-2的脂质包膜主要由甘油磷脂组成,缺乏胆固醇和鞘脂,这似乎是其易感性的关键因素,将其与流感病毒区分开来,后者的脂质谱中含有更丰富的抗氧化应激成分。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Analysis of the SARS-CoV-2 inactivation mechanism using violet-blue light (405 nm).

The study evaluated the effects of violet-blue light (VBL) on cell viability and replication, carbonylation of three structural proteins (S, E, and N) and one non-structural protein (NSP13), and direct damage to the RNA of SARS-CoV-2. The virus was exposed to increasing doses of VBL along with influenza A and B viruses to compare their susceptibility. At the highest dose (21.6 J/cm2), SARS-CoV-2 was significantly more susceptible to VBL than the influenza viruses, with a reduction in viral titer of 2.33 log10. Viral RNA did not show significant changes after exposure to VBL, as demonstrated by next-generation sequencing and real-time PCR quantification, suggesting that the inactivation process does not involve direct nucleic acid damage. To exclude the role of the culture suspension in the inactivation process, virus viability experiments were performed using different dilutions of Dulbecco's modified Eagle's medium (DMEM) in phosphate-buffered saline (PBS). The results indicated that the suspension medium played a secondary role in virus inactivation, as viability did not increase with increasing DMEM dilution. Subsequent tests with three different antioxidants (NAC, AsA, and SOD) at different concentrations prevented viral inactivation, from 99.99% to 85.43% (with SOD 0.003 mM). Carbonylation of S and E proteins was more pronounced when viruses were suspended in DMEM rather than PBS, although the tests demonstrated that the intrinsic properties of the viral membrane were a crucial element to consider in relation to its susceptibility to VBL.IMPORTANCELight-based disinfection methods are often used in combination with other cleaning methods due to their non-invasive nature, versatility, and environmental benefits. VBL is an effective approach as it induces the production of reactive oxygen species that reduce microbial viability. In this study, lipid peroxidation was identified as an important factor affecting the structural integrity and function of the viral envelope, reducing its ability to interact with host cells and consequently its ability to be infectious. The lipid envelope of SARS-CoV-2, composed mainly of glycerophospholipids and lacking cholesterol and sphingolipids, appears to be the critical factor in its susceptibility, distinguishing it from influenza viruses, which have a lipid profile richer in components that protect against oxidative stress.

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来源期刊
Applied and Environmental Microbiology
Applied and Environmental Microbiology 生物-生物工程与应用微生物
CiteScore
7.70
自引率
2.30%
发文量
730
审稿时长
1.9 months
期刊介绍: Applied and Environmental Microbiology (AEM) publishes papers that make significant contributions to (a) applied microbiology, including biotechnology, protein engineering, bioremediation, and food microbiology, (b) microbial ecology, including environmental, organismic, and genomic microbiology, and (c) interdisciplinary microbiology, including invertebrate microbiology, plant microbiology, aquatic microbiology, and geomicrobiology.
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