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 × 10 5 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 × 10 4 TCID 50 /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 × 10 3 TCID 50 /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.
【저자키워드】 SARS-CoV-2, biosafety, PCR, high throughput, Clinical diagnosis, laboratory-acquired infection, 【초록키워드】 Treatment, Efficacy, diagnostic, virus, Laboratory, RNA extraction, clinical samples, Culture, VTM, lysis buffer, inactivation, Rapid, platform, Combination, infectious dose, reduction, Volume, Viral transport medium, tissue, titre, isopropanol, Qiagen, acid, heat, operator, Thermo Fisher, TCID, viral RNA extraction, guanidinium, Result, shown, tested, addition, evaluated, reduced, demonstrated, 【제목키워드】 RNA, extraction, the SARS-CoV-2,