| Autor: |
Dominic Wooding, Kate Buist, Alessandra Romero-Ramirez, Helen Savage, Rachel Watkins, Daisy Bengey, Caitlin Greenland-Bews, Caitlin R. Thompson, Nadia Kontogianni, Richard Body, Gail Hayward, Rachel L. Byrne, Susan Gould, Christopher Myerscough, Barry Atkinson, Victoria Shaw, Bill Greenhalf, Emily Adams, Ana Cubas-Atienzar, Saye Khoo, Tom Fletcher, Thomas Edwards |
| Jazyk: |
angličtina |
| Rok vydání: |
2024 |
| Předmět: |
|
| Zdroj: |
mSphere, Vol 9, Iss 11 (2024) |
| Druh dokumentu: |
article |
| ISSN: |
2379-5042 |
| DOI: |
10.1128/msphere.00304-24 |
| Popis: |
ABSTRACT 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 in combination with RT-qPCR of culture supernatants from the first passage provides the greatest overall detection of Delta viral replication (22 of 32, 68.8%), being able to identify viable virus in 83.3% (20 of 24) of clinical samples with initial Ct values of |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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