University of Veterinary Medicine Hannover, Hannover, Germany.
Vet Pathol. 2024 Mar;61(2):201-206. doi: 10.1177/03009858231197128. Epub 2023 Sep 12.
The SARS-CoV-2 pandemic required the immediate need to transfer inactivated tissue from biosafety level (BSL)-3 to BSL-1 areas to enable downstream analytical methods. No validated SARS-CoV-2 inactivation protocols were available for either formaldehyde (FA)-fixed or glutaraldehyde (GA)-fixed tissues. Therefore, representative tissue from ferrets and hamsters was spiked with 2.2 × 10 tissue culture infectious dose 50% per ml (TCID/ml) SARS-CoV-2 or were obtained from mice experimentally infected with SARS-CoV-2. SARS-CoV-2 inactivation was demonstrated with 4% FA or 5% GA at room temperature for 72 hours by a titer reduction of up to 10 TCID/ml in different animal tissues with a maximum protein content of 100 µg/mg and a thickness of up to 10 mm for FA and 8 mm for GA. Our protocols can be easily adapted for validating the inactivation of other pathogens to allow for the transfer of biological samples from BSL-3 areas to BSL-1 laboratories.
SARS-CoV-2 大流行需要立即将来自生物安全级别 3(BSL-3)的灭活组织转移到 BSL-1 区域,以启用下游分析方法。甲醛(FA)固定或戊二醛(GA)固定组织均没有经过验证的 SARS-CoV-2 灭活方案。因此,从雪貂和仓鼠中提取了代表性组织,用每毫升 2.2×10 组织培养感染剂量 50%(TCID/ml)的 SARS-CoV-2 进行接种,或从感染 SARS-CoV-2 的实验小鼠中获得。通过在室温下用 4%FA 或 5%GA 处理 72 小时,在不同动物组织中,病毒滴度降低了 10 TCID/ml,最大蛋白含量为 100 µg/mg,厚度为 10mm(FA)和 8mm(GA),可以证明 SARS-CoV-2 的灭活。我们的方案可以轻松适应其他病原体灭活的验证,以允许将生物样本从 BSL-3 区域转移到 BSL-1 实验室。