Hume Adam J, Ames Joshua, Rennick Linda J, Duprex W Paul, Marzi Andrea, Tonkiss John, Mühlberger Elke
Department of Microbiology, School of Medicine, Boston University, 620 Albany Street, Boston, MA 02118, USA.
National Emerging Infectious Diseases Laboratories (NEIDL), Boston University, 620 Albany Street, Boston, MA 02118, USA.
Viruses. 2016 Jul 22;8(7):204. doi: 10.3390/v8070204.
Effective inactivation of biosafety level 4 (BSL-4) pathogens is vital in order to study these agents safely. Gamma irradiation is a commonly used method for the inactivation of BSL-4 viruses, which among other advantages, facilitates the study of inactivated yet morphologically intact virions. The reported values for susceptibility of viruses to inactivation by gamma irradiation are sometimes inconsistent, likely due to differences in experimental protocols. We analyzed the effects of common sample attributes on the inactivation of a recombinant vesicular stomatitis virus expressing the Zaire ebolavirus glycoprotein and green fluorescent protein. Using this surrogate virus, we found that sample volume and protein content of the sample modulated viral inactivation by gamma irradiation but that air volume within the sample container and the addition of external disinfectant surrounding the sample did not. These data identify several factors which alter viral susceptibility to inactivation and highlight the usefulness of lower biosafety level surrogate viruses for such studies. Our results underscore the need to validate inactivation protocols of BSL-4 pathogens using "worst-case scenario" procedures to ensure complete sample inactivation.
有效灭活生物安全4级（BSL-4）病原体对于安全研究这些病原体至关重要。伽马辐射是一种常用的灭活BSL-4病毒的方法，它具有诸多优点，包括有助于对已灭活但形态完整的病毒粒子进行研究。报道的病毒对伽马辐射灭活的敏感性数值有时并不一致，这可能是由于实验方案的差异所致。我们分析了常见样本属性对表达扎伊尔埃博拉病毒糖蛋白和绿色荧光蛋白的重组水疱性口炎病毒灭活的影响。使用这种替代病毒，我们发现样本体积和样本中的蛋白质含量会调节伽马辐射对病毒的灭活作用，但样本容器内的空气体积以及样本周围添加外部消毒剂则不会。这些数据确定了几个改变病毒对灭活敏感性的因素，并突出了较低生物安全水平替代病毒在此类研究中的有用性。我们的结果强调需要使用“最坏情况”程序来验证BSL-4病原体的灭活方案，以确保样本完全灭活。
