biostudies-literatureWooding DWellcome Truste0030424https://www.ebi.ac.uk/biostudies/studies/S-EPMC11580409biostudies-literatureUnknown9(11)Clinical trials of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) therapeutics often include virological secondary endpoints to compare viral clearance and viral load reduction between treatment and placebo arms. This is typically achieved using quantitative reverse-transcriptase PCR (RT-qPCR), which cannot differentiate replicant competent virus from non-viable virus or free RNA, limiting its utility as an endpoint. Culture-based methods for SARS-CoV-2 exist; however, these are often insensitive and poorly standardized for use as clinical trial endpoints. We report optimization of a culture-based approach evaluating three cell lines, three detection methods, and key culture parameters. We show that Vero-angiotensin-converting enzyme 2-transmembrane serine protease 2 cells inmSphereOptimization of SARS-CoV-2 culture from clinical samples for clinical trial applications.PMC11580409221590/Z/20/ZWooding DDavis KTate VNicholson BBody RLendrem CLogan MHayward GCONDOR Steering GroupEdwards TWatkins RFletcher TJones NJones WSavage HLewington AAllen AJMyerscough CAtkinson BRiley BBengey DBuckle PLee JThompson CRMicocci MCubas-Atienzar ALasserson DPerera-Salazar RDark PSimpson AJWilcox MReynard CTurner PBuist KHarden CGreenhalf BHalstead APrice DAShaw VAdams EByrne RLZhifang MKontogianni NInkson CPrestwich GRomero-Ramirez AKhoo SGreenland-Bews CGordon AGould SBraybrook JfalseOptimization of SARS-CoV-2 culture from clinical samples for clinical trial applications.Clinical trials of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) therapeutics often include virological secondary endpoints to compare viral clearance and viral load reduction between treatment and placebo arms. This is typically achieved using quantitative reverse-transcriptase PCR (RT-qPCR), which cannot differentiate replicant competent virus from non-viable virus or free RNA, limiting its utility as an endpoint. Culture-based methods for SARS-CoV-2 exist; however, these are often insensitive and poorly standardized for use as clinical trial endpoints. We report optimization of a culture-based approach evaluating three cell lines, three detection methods, and key culture parameters. We show that Vero-angiotensin-converting enzyme 2-transmembrane serine protease 2 cells in2024-01-01T00:00:00Z2024 Nov2025-04-04T13:40:19.429Z2025-04-04T13:40:19.429ZS-EPMC115804093941228310.1128/msphere.00304-24