Tanaka Tsuyoshi, Nogariya Osamu, Shionoiri Nozomi, Maeda Yoshiaki, Arakaki Atsushi
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
J Biosci Bioeng. 2018 Jul;126(1):63-68. doi: 10.1016/j.jbiosc.2018.01.018. Epub 2018 Feb 25.
UV-C treatment has been shown to be a powerful way to inactivate non-enveloped viruses in water samples. However, little is known about how the viruses were inactivated by UV-C radiation. In this study, we investigated the inactivation mechanism of a single-stranded RNA (ssRNA) non-enveloped virus, feline calicivirus (FCV), as a surrogate for the human norovirus, using UV-C radiation with different wavelengths. Integrated molecular analyses using RT-qPCR, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and mass spectrometry were employed to evaluate the extent of ssRNA genome and protein degradation. UV-C radiation of FCV efficiently impaired the infectivity of FCV in mammalian cells. We also identified degradation of the RNA genome, whose copy numbers decreased from 48% to 56% following UV or UV radiation. Significant degradation of capsid protein was not observed, whereas oxidation of amino acid residues in the major capsid protein VP-1 was determined. Our results suggest that damage to the RNA genome is primarily responsible for the observed decrease in FCV infectivity of CRFK cells. This study provides not only relevant baseline data but also an overview and possible mechanism for the disinfection of non-enveloped ssRNA viruses using UV-C radiation.
紫外线C（UV-C）处理已被证明是一种使水样中非包膜病毒失活的有效方法。然而，对于紫外线C辐射如何使病毒失活却知之甚少。在本研究中，我们以猫杯状病毒（FCV）作为人诺如病毒的替代物，使用不同波长的紫外线C辐射，研究了单链RNA（ssRNA）非包膜病毒的失活机制。采用实时定量聚合酶链反应（RT-qPCR）、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和质谱等综合分子分析方法，评估ssRNA基因组和蛋白质的降解程度。FCV的紫外线C辐射有效损害了FCV在哺乳动物细胞中的感染性。我们还鉴定出RNA基因组的降解，紫外线或紫外线辐射后其拷贝数从48%降至56%。未观察到衣壳蛋白的显著降解，但确定了主要衣壳蛋白VP-1中氨基酸残基的氧化。我们的结果表明，RNA基因组的损伤是导致观察到的FCV对CRFK细胞感染性下降的主要原因。本研究不仅提供了相关的基线数据，还概述了使用紫外线C辐射对非包膜ssRNA病毒进行消毒的可能机制。
