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Rotavirus Virus-Like Particles as Surrogates in Environmental Persistence and Inactivation Studies

机译：轮状病毒样颗粒在环境持久性和灭活研究中作为替代物

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Virus-like particles (VLPs) with the full-length VP2 and VP6 rotavirus capsid proteins, produced in the baculovirus expression system, have been evaluated as surrogates of human rotavirus in different environmental scenarios. Green fluorescent protein-labeled VLPs (GFP-VLPs) and particles enclosing a heterologous RNA (pseudoviruses), whose stability may be monitored by flow cytometry and antigen capture reverse transcription-PCR, respectively, were used. After 1 month in seawater at 20°C, no significant differences were observed between the behaviors of GFP-VLPs and of infectious rotavirus, whereas pseudovirus particles showed a higher decay rate. In the presence of 1 mg of free chlorine (FC)/liter both tracers persisted longer in freshwater at 20°C than infectious viruses, whereas in the presence of 0.2 mg of FC/liter no differences were observed between tracers and infectious rotavirus at short contact times. However, from 30 min of contact with FC onward, the decay of infectious rotavirus was higher than that of recombinant particles. The predicted Ct value for a 90% reduction of GFP-VLPs or pseudoviruses induces a 99.99% inactivation of infectious rotavirus. Both tracers were more resistant to UV light irradiation than infectious rotavirus in fresh and marine water. The effect of UV exposure was more pronounced on pseudovirus than in GFP-VLPs. In all types of water, the UV dose to induce a 90% reduction of pseudovirus ensures a 99.99% inactivation of infectious rotavirus. Recombinant virus surrogates open new possibilities for the systematic validation of virus removal practices in actual field situations where pathogenic agents cannot be introduced.
机译：杆状病毒表达系统中产生的带有全长VP2和VP6轮状病毒衣壳蛋白的病毒样颗粒（VLP）已被评估为人类轮状病毒在不同环境中的替代物。使用了绿色荧光蛋白标记的VLP（GFP-VLP）和包裹异源RNA的颗粒（假病毒），其稳定性可以分别通过流式细胞仪和抗原捕获逆转录PCR进行监测。在20°C的海水中放置1个月后，GFP-VLP和感染性轮状病毒的行为之间没有发现显着差异，而假病毒颗粒显示出更高的衰减率。在存在1 mg游离氯（FC）/升的情况下，两种示踪剂在20°C的淡水中比感染病毒的持续时间更长，而在存在0.2 mg FC /升的示踪剂时，示踪剂和传染性轮状病毒之间没有观察到差异联系时间。然而，从与FC接触30分钟开始，感染性轮状病毒的衰减高于重组颗粒。 GFP-VLP或假病毒减少90％的预计Ct值会导致99.99％的感染性轮状病毒灭活。与淡水和海水中的感染性轮状病毒相比，这两种示踪剂对紫外线的抵抗力都更高。紫外线对假病毒的影响比GFP-VLP更明显。在所有类型的水中，紫外线剂量可诱导伪病毒减少90％，可确保99.99％的感染性轮状病毒灭活。重组病毒替代品为在无法引入病原体的实际现场情况下系统验证病毒去除实践提供了新的可能性。

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