Liew Oi Wah, Fanusi Felic, Ng Jessica Yan Xia, Ahidjo Bintou Ahmadou, Ling Samantha Shi Min, Lilyanna Shera, Chong Jenny Pek Ching, Lim Angeline Eng Siew, Lim Wei Zheng, Ravindran Sindhu, Chu Justin Jang Hann, Lim Shir Lynn, Richards Arthur Mark
Cardiovascular Research Institute, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, 14 Medical Drive, Singapore 117599, Singapore.
NUS Medicine BSL3 Core Facility, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, 14 Medical Drive, Singapore 117599, Singapore.
ACS Omega. 2022 Jul 14;7(29):25510-25520. doi: 10.1021/acsomega.2c02585. eCollection 2022 Jul 26.
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) inactivation is an important step toward enhanced biosafety in testing facilities and affords a reduction in the biocontainment level necessary for handling virus-positive biological specimens. Virus inactivation methods commonly employ heat, detergents, or combinations thereof. In this work, we address the dearth of information on the efficacy of SARS-CoV-2 inactivation procedures in plasma and their downstream impact on immunoassays. We evaluated the effects of heat (56 °C for 30 min), detergent (1-5% Triton X-100), and solvent-detergent (SD) combinations [0.3-1% tri--butyl phosphate (TNBP) and 1-2% Triton X-100] on 19 immunoassays across different assay formats. Treatments are deemed immunoassay-compatible when the average and range of percentage recovery (treated concentration relative to untreated concentration) lie between 90-110 and 80-120%, respectively. We show that SD treatment (0.3% TNBP/1% Triton-X100) is compatible with more than half of the downstream immunoassays tested and is effective in reducing SARS-CoV-2 infectivity in plasma to below detectable levels in plaque assays. This facile method offers enhanced safety for laboratory workers handling biological specimens in clinical and research settings.
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)灭活是提高检测设施生物安全性的重要一步,并且可以降低处理病毒阳性生物样本所需的生物安全水平。病毒灭活方法通常采用加热、洗涤剂或两者结合的方式。在这项研究中,我们解决了SARS-CoV-2在血浆中的灭活程序效果及其对免疫测定的下游影响方面信息匮乏的问题。我们评估了加热(56℃ 30分钟)、洗涤剂(1-5% Triton X-100)以及溶剂-洗涤剂(SD)组合[0.3-1%磷酸三丁酯(TNBP)和1-2% Triton X-100]对不同检测形式的19种免疫测定的影响。当回收率百分比(处理后浓度相对于未处理浓度)的平均值和范围分别在90-110%和80-120%之间时,这些处理被认为与免疫测定兼容。我们表明,SD处理(0.3% TNBP/1% Triton-X100)与超过一半的测试下游免疫测定兼容,并且在噬斑测定中能有效将血浆中的SARS-CoV-2感染性降低到可检测水平以下。这种简便的方法为临床和研究环境中处理生物样本的实验室工作人员提供了更高的安全性。