Validation of the Thermo Scientific™ SARS-CoV-2 RT-PCR Detection Workflow for the Detection of SARS-CoV-2 from Stainless Steel Environmental Surface Swabs AOAC Performance Tested MethodSM 012103.

Background The Thermo Scientific™ SARS-CoV-2 RT-PCR Detection Workflow, packaged with Applied Biosystems™ TaqMan™ 2019-nCoV Assay Kit v1 targets three different SARS-CoV-2 genomic regions in a single RT-PCR reaction. Objective To validate the Thermo Scientific™ SARS-CoV-2 RT-PCR Workflow, for the detection of SARS-CoV-2 virus on stainless steel surfaces as part of the AOAC Performance Tested Method SM Emergency Response Validation program. Methods The Applied Biosystems™ TaqMan™ 2019-nCoV Assay Kit v1, as part of the Thermo Scientific SARS-CoV-2 RT-PCR Workflow was evaluated for specificity using in silico analysis of 15,764 SARS-CoV-2 sequences and 65 exclusivity organisms. The Thermo Scientific SARS-CoV-2 RT-PCR Workflow was evaluated in an unpaired study for one environmental surface (stainless steel) and compared to the U.S. Centers for Disease Control and Prevention 2019-Novel Coronavirus RT-PCR Diagnostic Panel, Instructions for Use (Revision 4, Effective 6/12/2020). Results In silico analysis showed that of the 15,756 target SARS-CoV-2 genomes analyzed, 99% of the strains/isolates are perfectly matched to at least two of the three assays, and more than 90% have 100% homology to all three assays (ORF1ab, N-gene, S-gene) in the SARS-CoV-2 Kit. None of the 65 non-target strain genomes analyzed showed matching sequences. In the matrix study, the Thermo Scientific SARS-CoV-2 workflow showed comparable detection to the CDC method. Conclusions The Thermo Scientific SARS-CoV-2 RT-PCR Workflow is an effective procedure for detection of RNA from SARS-CoV-2 virus from stainless steel. Highlights The workflow provides equivalent performance results with the two tested RNA extraction platforms and the two tested RT-PCR Instruments.