National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB R3E 3R2, Canada.
Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, MB R3E 0J9, Canada.
Viruses. 2024 Aug 24;16(9):1354. doi: 10.3390/v16091354.
High-consequence pathogens such as the Ebola, Marburg, and Lassa viruses are handled in maximum-containment biosafety level 4 (BSL-4) laboratories. Genetic material is often isolated from such viruses and subsequently removed from BSL-4 laboratories for a multitude of downstream analyses using readily accessible technologies and equipment available at lower-biosafety level laboratories. However, it is essential to ensure that these materials are free of viable viruses before removal from BSL-4 laboratories to guarantee sample safety. This study details the in-house procedure used for validating the inactivation of Ebola, Marburg, and Lassa virus cultures after incubation with AVL lysis buffer (Qiagen) and ethanol. This study's findings show that no viable virus was detectable when high-titer cultures of Ebola, Marburg, and Lassa viruses were incubated with AVL lysis buffer for 10 min, followed by an equal volume of 95% ethanol for 3 min, using a method with a sensitivity of ≤0.8 log TCID as the limit of detection.
高致病性病原体，如埃博拉、马尔堡和拉萨热病毒，在最高防护生物安全级别 4（BSL-4）实验室中进行处理。这些病毒的遗传物质通常从实验室中分离出来，然后转移到 BSL-4 实验室之外，以便在更低生物安全级别的实验室中使用易于获取的技术和设备进行多种下游分析。然而，在将这些材料从 BSL-4 实验室中移除之前，必须确保它们不含有活性的病毒，以保证样品的安全。本研究详细介绍了一种内部程序，用于验证在与 AVL 裂解缓冲液（Qiagen）和乙醇孵育后，埃博拉、马尔堡和拉萨热病毒培养物的失活情况。本研究的结果表明，当高滴度的埃博拉、马尔堡和拉萨热病毒培养物用 AVL 裂解缓冲液孵育 10 分钟，然后再用等体积的 95%乙醇孵育 3 分钟时，使用灵敏度≤0.8 log TCID 作为检测限的方法，无法检测到有活性的病毒。
