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Rapid luminescence-based screening method for SARS- CoV-2 inhibitors discovery

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Názov: Rapid luminescence-based screening method for SARS- CoV-2 inhibitors discovery
Autori: Abdeldjalil Madani, Nadine Alvarez, Steven Park, Madhuvika Murugan, David S. Perlin
Zdroj: SLAS Discovery, Vol 31, Iss , Pp 100211- (2025)
Informácie o vydavateľovi: Elsevier, 2025.
Rok vydania: 2025
Zbierka: LCC:Medicine (General)
LCC:Biotechnology
Predmety: Drug screening, HTS, Viruses, Antivirals, Nano-luciferase, Bsl-3 viruses, Medicine (General), R5-920, Biotechnology, TP248.13-248.65
Popis: The COVID-19 pandemic has emphasized the necessity for rapid and adaptable drug screening platforms against live pathogenic viruses that require high levels of biosafety containment. Conventional antiviral testing is time-consuming and labor-intensive. Here, we outline the design and validation of a semi-automated drug-screening platform for SARS-CoV-2 that utilizes multiple liquid handlers, a stable A549 cell line expressing ACE2 and TMPRSS2 receptors, and a recombinant SARS-CoV-2 strain harboring the nano-luciferase gene. This platform allows for accelerated low-, mid-, and high-throughput screenings by bypassing the virus inactivation and the staining steps compared to assays utilizing fluorescent reporter viruses or immunofluorescence. First, we demonstrated that the luminescence signal obtained at 24 h post-infection is robust and can be used as a surrogate for fluorescent reporter viruses and immunofluorescence assays that require 48 h incubation post infection. We confirmed the susceptibility of the reporter virus to a panel of reference drugs and validated the luminescence signal in 96- and 384-well plates in accordance with NIH criteria for high-throughput screening. The validation assays showed reproducible results, robust Z factor of ≥0.5, and a coefficient of variation of
Druh dokumentu: article
Popis súboru: electronic resource
Jazyk: English
ISSN: 2472-5552
Relation: http://www.sciencedirect.com/science/article/pii/S2472555225000048; https://doaj.org/toc/2472-5552
DOI: 10.1016/j.slasd.2025.100211
Prístupová URL adresa: https://doaj.org/article/3318a48fecf74e2e92ff1d68c70c653c
Prístupové číslo: edsdoj.3318a48fecf74e2e92ff1d68c70c653c
Databáza: Directory of Open Access Journals
Popis
Abstrakt:The COVID-19 pandemic has emphasized the necessity for rapid and adaptable drug screening platforms against live pathogenic viruses that require high levels of biosafety containment. Conventional antiviral testing is time-consuming and labor-intensive. Here, we outline the design and validation of a semi-automated drug-screening platform for SARS-CoV-2 that utilizes multiple liquid handlers, a stable A549 cell line expressing ACE2 and TMPRSS2 receptors, and a recombinant SARS-CoV-2 strain harboring the nano-luciferase gene. This platform allows for accelerated low-, mid-, and high-throughput screenings by bypassing the virus inactivation and the staining steps compared to assays utilizing fluorescent reporter viruses or immunofluorescence. First, we demonstrated that the luminescence signal obtained at 24 h post-infection is robust and can be used as a surrogate for fluorescent reporter viruses and immunofluorescence assays that require 48 h incubation post infection. We confirmed the susceptibility of the reporter virus to a panel of reference drugs and validated the luminescence signal in 96- and 384-well plates in accordance with NIH criteria for high-throughput screening. The validation assays showed reproducible results, robust Z factor of ≥0.5, and a coefficient of variation of
ISSN:24725552
DOI:10.1016/j.slasd.2025.100211