Molecular Virology, Department Biomedicine, University of Basel, 4009 Basel, Switzerland.
Viruses. 2023 Aug 4;15(8):1693. doi: 10.3390/v15081693.
Numerous mammalian viruses are routinely analyzed in clinical diagnostic laboratories around the globe or serve as indispensable model systems in viral research. Potentially infectious viral entities are handled as blood, biopsies, or cell and tissue culture samples. Countless protocols describe methods for virus fixation and inactivation, yet for many, a formal proof of safety and completeness of inactivation remains to be shown. While modern nucleic acid extraction methods work quite effectively, data are largely lacking on possible residual viral infectivity, e.g., when assessed after extended culture times, which maximizes the sensitivity for low levels of residual infectiousness. Therefore, we examined the potency and completeness of inactivation procedures on virus-containing specimens when applying commonly used fixatives like formaldehyde or nucleic acid extraction/lysis buffers. Typical representatives of different virus classes, including RNA and DNA viruses, enveloped and non-enveloped, such as adenovirus, enterovirus, lentivirus, and coronavirus, were used, and the reduction in the in vitro infectiousness was assessed for standard protocols. Overall, a 30-minute incubation with formaldehyde at room temperature effectively inactivated all tested enveloped and non-enveloped viruses. Full inactivation of HIV-1 and ECHO-11 was also achieved with all buffers in the test, whereas for SARS-CoV-2 and AdV-5, only five of the seven lysis buffers were fully effective under the tested conditions.
全球范围内,许多哺乳动物病毒在临床诊断实验室中被常规分析,或作为病毒研究中不可或缺的模型系统。潜在传染性的病毒实体作为血液、活检、细胞和组织培养样本进行处理。无数方案描述了病毒固定和失活的方法,但对于许多方法来说,仍然需要证明其安全性和失活的完整性。虽然现代核酸提取方法非常有效,但关于可能存在的残留病毒感染性的数据在很大程度上仍然缺乏,例如,在延长的培养时间后进行评估时,这最大限度地提高了低水平残留传染性的敏感性。因此,我们研究了当应用常用固定剂(如甲醛或核酸提取/裂解缓冲液)时,含病毒标本中灭活程序的效力和完整性。使用了不同病毒类别的典型代表,包括 RNA 和 DNA 病毒、包膜和非包膜病毒,如腺病毒、肠道病毒、慢病毒和冠状病毒,并评估了标准方案中体外传染性的降低情况。总体而言,室温下 30 分钟孵育甲醛可有效灭活所有测试的包膜和非包膜病毒。所有测试缓冲液均能完全灭活 HIV-1 和 ECHO-11,而对于 SARS-CoV-2 和 AdV-5,只有七种裂解缓冲液中的五种在测试条件下完全有效。