Optimal preparation of SARS-CoV-2 viral transport medium for culture

Abstract Introduction The sudden arrival of the COVID-19 pandemic placed significant stresses on supply chains including viral transport medium (VTM). VTM that was urgently required needed to support replication, as well other routine diagnostic approaches. We describe preparation and validation testing for rapidly expanding testing, where capacity preserve integrity, culture, isolation full sequence analysis, maintained. Methods prepared using different methods sterilization then ‘spiked’ with virus. investigated culture in Vero cells, nucleic acid detection by quantitative PCR. Results best results were obtained filter autoclave-based sterilization. proved robust culture-based analyses provided inoculated stored at 4 °C, tested within 48 h. filtered also supported PCR-based diagnosis least 5 days when mock held room temperature. Discussion manual handling production, filling sterilization, optimized. SARS-CoV-2 spiked into assess measure effects storage time temperature upon performance. While most protocols will not require replication competent virus, use high quality allow next phase laboratory analysis pandemic, drug antibody susceptibility re-isolated SARS-CoV-2, vaccine escape mutants.