Rapid and demonstrable inactivation of SARS-CoV-2 is crucial to ensure operator safety during high-throughput testing of clinical samples. The inactivation efficacy of SARS-CoV-2 was evaluated using commercially available lysis buffers from three viral RNA extraction kits used on two high-throughput (96-well) RNA extraction platforms (Qiagen QIAcube HT and the Thermo Fisher KingFisher Flex) in combination with thermal treatment. Buffer volumes and sample ratios were chosen for their optimised suitability for RNA extraction rather than inactivation efficacy and tested against a representative sample type: SARS-CoV-2 spiked into viral transport medium (VTM). A lysis buffer mix from the MagMAX Pathogen RNA/DNA kit (Thermo Fisher), used on the KingFisher Flex, which included guanidinium isothiocyanate (GITC), a detergent, and isopropanol, demonstrated a minimum inactivation efficacy of 1 × 10⁵ tissue culture infectious dose (TCID)₅₀/ml. Alternative lysis buffer mixes from the MagMAX Viral/Pathogen Nucleic Acid kit (Thermo Fisher) also used on the KingFisher Flex and from the QIAamp 96 Virus QIAcube HT Kit (Qiagen) used on the QIAcube HT (both of which contained GITC and a detergent) reduced titres by 1 × 10⁴ TCID₅₀/ml but did not completely inactivate the virus. Heat treatment alone (15 min, 68°C) did not completely inactivate the virus, demonstrating a reduction of 1 × 10³ TCID₅₀/ml. When inactivation methods included both heat treatment and addition of lysis buffer, all methods were shown to completely inactivate SARS-CoV-2 inactivation against the viral titres tested. Results are discussed in the context of the operation of a high-throughput diagnostic laboratory.

SARS-CoV-2 的快速且可证明的灭活对于确保临床样本高通量测试期间操作员的安全至关重要。使用来自两种高​​通量（96 孔）RNA 提取平台（Qiagen QIAcube HT 和 Thermo Fisher KingFisher Flex）上使用的三种病毒 RNA 提取试剂盒的市售裂解缓冲液评估 SARS-CoV-2 的灭活功效。热处理。选择缓冲液体积和样品比例是因为它们优化了 RNA 提取的适用性，而不是灭活功效，并针对代表性样品类型进行了测试：SARS-CoV-2 掺入病毒转运介质 (VTM)。来自 MagMAX Pathogen RNA/DNA 试剂盒 (Thermo Fisher) 的裂解缓冲液混合物，用于 KingFisher Flex，其中包括异硫氰酸胍 (GITC)、去污剂和异丙醇，⁵组织培养感染剂量（TCID）₅₀ /ml。来自同样用于 KingFisher Flex 的 MagMAX 病毒/病原体核酸试剂盒 (Thermo Fisher) 和来自用于 QIAcube HT 的 QIAamp 96 病毒 QIAcube HT 试剂盒 (Qiagen) 的替代裂解缓冲液混合物（均含有 GITC 和去污剂）滴度降低了 1 × 10 ⁴ TCID ₅₀ /ml，但并未完全灭活病毒。单独热处理（15 分钟，68°C）并未完全灭活病毒，证明降低了 1 × 10 ³ TCID ₅₀/毫升。当灭活方法包括热处理和添加裂解缓冲液时，所有方法都显示针对测试的病毒滴度完全灭活 SARS-CoV-2。结果在高通量诊断实验室运行的背景下进行讨论。