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However, antiviral combinations may be more potent and lead to faster viral clearance, but the methods for screening antiviral combinations against respiratory viruses are not well established and labor-intensive. Here, we describe a time-efficient (72\u201396 h) and simple\n                      <jats:italic>in vitro<\/jats:italic>\n                      drug-sensitivity assay for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using standard 96-well plates. We employ different synergy models (zero interaction potency, highest single agent, Loewe, Bliss) to determine the efficacy of antiviral therapies and synergistic combinations against ancestral and emerging clinical SARS-CoV-2 strains. We found that monotherapy of remdesivir, nirmatrelvir, and active metabolite of molnupiravir (EIDD-1931) demonstrated baseline EC50s within clinically achievable levels of 4.34 mg\/L (CI: 3.74\u20134.94 mg\/L), 1.25 mg\/L (CI: 1.10\u20131.45 mg\/L), and 0.25 mg\/L (CI: 0.20\u20130.30 mg\/L), respectively, against the ancestral SARS-CoV-2 strain. However, their efficacy varied against newer Omicron variants BA.1.1.15 and BA.2, particularly with the protease inhibitor nirmatrelvir. We also found that remdesivir and nirmatrelvir have a consistent, strong synergistic effect (Bliss synergy score &gt;10) at clinically relevant drug concentrations (nirmatrelvir 0.25\u20131 mg\/L with remdesivir 1\u20134 mg\/L) across all SARS-CoV-2 strains tested. This method offers a practical tool that streamlines the identification of effective combination therapies and the detection of antiviral resistance. Our findings support the use of antiviral drug combinations targeting multiple viral components to enhance COVID-19 treatment efficacy, particularly in the context of emerging viral strains.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.01233-24","type":"journal-article","created":{"date-parts":[[2024,12,17]],"date-time":"2024-12-17T09:54:03Z","timestamp":1734429243000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Efficient\n                    <i>in vitro<\/i>\n                    assay for evaluating drug efficacy and synergy against emerging SARS-CoV-2 strains"],"prefix":"10.1128","volume":"69","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4511-9171","authenticated-orcid":true,"given":"Maximillian","family":"Woodall","sequence":"first","affiliation":[{"name":"UCL Great Ormond Street Institute of Child Health","place":["London, United 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