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High-Throughput SARS-CoV-2 Antiviral Testing Method Using the Celigo Image Cytometer.
St Clair, Laura A; Chan, Leo Li-Ying; Boretsky, Adam; Lin, Bo; Spedding, Michael; Perera, Rushika.
Afiliación
  • St Clair LA; Center for Vector-borne Infectious Diseases, Department of Microbiology, Immunology, and Pathology, Colorado State University, CO, 80523, Fort Collins, USA.
  • Chan LL; Center for Metabolism of Infectious Diseases (C4MInD), Colorado State University, 3185 Rampart Rd, Fort Collins, CO, 80523, USA.
  • Boretsky A; Department of Advanced Technology R&D, Revvity, 360 Merrimack St. Suite 200, Lawrence, MA, 01843, USA. leo.chan@perkinelmer.com.
  • Lin B; Department of Advanced Technology R&D, Revvity, 360 Merrimack St. Suite 200, Lawrence, MA, 01843, USA.
  • Spedding M; Department of Advanced Technology R&D, Revvity, 360 Merrimack St. Suite 200, Lawrence, MA, 01843, USA.
  • Perera R; Spedding Research Solutions SAS, Le Vesinet, France.
J Fluoresc ; 34(2): 561-570, 2024 Mar.
Article en En | MEDLINE | ID: mdl-37310590
The COVID-19 pandemic has created a worldwide public health crisis that has since resulted in 6.8 million reported deaths. The pandemic prompted the immediate response of researchers around the world to engage in rapid vaccine development, surveillance programs, and antiviral testing, which resulted in the delivery of multiple vaccines and repurposed antiviral drug candidates. However, the emergence of new highly transmissible SARS-CoV-2 variants has renewed the desire for discovering new antiviral drug candidates with high efficacy against the emerging variants of concern. Traditional antiviral testing methods employ the plaque-reduction neutralization tests (PRNTs), plaque assays, or RT-PCR analysis, but each assay can be tedious and time-consuming, requiring 2-3 days to complete the initial antiviral assay in biologically relevant cells, and then 3-4 days to visualize and count plaques in Vero cells, or to complete cell extractions and PCR analysis. In recent years, plate-based image cytometers have demonstrated high-throughput vaccine screening methods, which can be adopted for screening potential antiviral drug candidates. In this work, we developed a high-throughput antiviral testing method employing the Celigo Image Cytometer to investigate the efficacy of antiviral drug candidates on SARS-CoV-2 infectivity using a fluorescent reporter virus and their safety by measuring the cytotoxicity effects on the healthy host cell line using fluorescent viability stains. Compared to traditional methods, the assays defined here eliminated on average 3-4 days from our standard processing time for antiviral testing. Moreover, we were able to utilize human cell lines directly that are not typically amenable to PRNT or plaque assays. The Celigo Image Cytometer can provide an efficient and robust method to rapidly identify potential antiviral drugs to effectively combat the rapidly spreading SARS-CoV-2 virus and its variants during the pandemic.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: SARS-CoV-2 / COVID-19 Límite: Animals / Humans Idioma: En Revista: J Fluoresc Asunto de la revista: BIOFISICA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: SARS-CoV-2 / COVID-19 Límite: Animals / Humans Idioma: En Revista: J Fluoresc Asunto de la revista: BIOFISICA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Países Bajos