Amodeo Davide, Marchi Serena, Fiaschi Lia, Raucci Luisa, Biba Camilla, Salvestroni Valentina, Trombetta Claudia Maria, Manini Ilaria, Zazzi Maurizio, Montomoli Emanuele, Vicenti Ilaria, Cevenini Gabriele, Messina Gabriele
Department of Medical Biotechnologies, University of Siena, Siena, Italy.
Department of Molecular and Developmental Medicine, University of Siena, Siena, Italy.
Appl Environ Microbiol. 2025 Jun 18;91(6):e0040325. doi: 10.1128/aem.00403-25. Epub 2025 May 14.
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/cm), SARS-CoV-2 was significantly more susceptible to VBL than the influenza viruses, with a reduction in viral titer of 2.33 log. 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.
该研究评估了蓝紫光（VBL）对细胞活力和复制、三种结构蛋白（S、E和N）和一种非结构蛋白（NSP13）的羰基化作用以及对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）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递增剂量的VBL中，以比较它们的敏感性。在最高剂量（21.6 J/cm）下，SARS-CoV-2对VBL的敏感性明显高于流感病毒，病毒滴度降低了2.33个对数。下一代测序和实时PCR定量结果表明，病毒RNA在暴露于VBL后未显示出显著变化，这表明灭活过程不涉及直接的核酸损伤。为了排除培养悬浮液在灭活过程中的作用，使用磷酸盐缓冲盐水（PBS）中不同稀释度的杜氏改良 Eagle培养基（DMEM）进行了病毒活力实验。结果表明，悬浮培养基在病毒灭活中起次要作用，因为随着DMEM稀释度的增加，病毒活力并未增加。随后用三种不同浓度的抗氧化剂（NAC、AsA和SOD）进行的测试阻止了病毒灭活，灭活率从99.99%降至85.43%（SOD浓度为0.003 mM时）。当病毒悬浮在DMEM中而不是PBS中时，S和E蛋白的羰基化更为明显，尽管测试表明病毒膜的固有特性是考虑其对VBL敏感性的关键因素。
重要性
基于光的消毒方法由于其非侵入性、多功能性和环境效益，常与其他清洁方法联合使用。VBL是一种有效的方法，因为它能诱导活性氧的产生，从而降低微生物的活力。在本研究中，脂质过氧化被确定为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，使其与流感病毒不同，流感病毒的脂质成分更丰富，含有能抵御氧化应激的成分。
