Optimisation of SARS-CoV-2 culture from clinical samples for clinical trial applications

Publication date: Mar 25, 2024

Clinical trials of 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 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 standardised for use as clinical trial endpoints. We report optimisation of a culture-based approach evaluating three cell lines, three detection methods, and key culture parameters. We show that Vero-ACE2-TMPRSS2 (VAT) cells in combination with RT-qPCR of culture supernatants from the first passage provides the greatest overall detection of Delta viral replication (22/32, 68.8%), being able to identify viable virus in 83.3% (20/24) of clinical samples with initial Ct values

Concepts Keywords
3manchester Based
Greenland Clinical
Pcr Cov
Virus Culture
Detection
Liverpool
Medrxiv
Passage
Positive
Preprint
Sars
Vat
Vero
Viral
Virus

Semantics

Type Source Name
disease VO viable
disease VO report
disease IDO cell
pathway KEGG Viral replication
drug DRUGBANK Corticorelin
disease VO frequency
disease MESH COVID 19
disease IDO susceptibility
disease MESH infection
disease IDO assay
disease VO volume

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