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Li, Li-Hsin; Chiu, Winston; Huang, Yun-An; Rasulova, Madina; Vercruysse, Thomas; Thibaut, Hendrik Jan; ter Horst, Sebastiaan; Rocha-Pereira, Joana; Vanhoof, Greet; Borrenberghs, Doortje; Goethals, Olivia; Kaptein, Suzanne J. F.; Leyssen, Pieter; Neyts, Johan; Dallmeier, Kai

Multiplexed multicolor antiviral assay amenable for high-throughput research

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  • Media type: E-Article
  • Title: Multiplexed multicolor antiviral assay amenable for high-throughput research
  • Contributor: Li, Li-Hsin; Chiu, Winston; Huang, Yun-An; Rasulova, Madina; Vercruysse, Thomas; Thibaut, Hendrik Jan; ter Horst, Sebastiaan; Rocha-Pereira, Joana; Vanhoof, Greet; Borrenberghs, Doortje; Goethals, Olivia; Kaptein, Suzanne J. F.; Leyssen, Pieter; Neyts, Johan; Dallmeier, Kai
  • Published: Springer Science and Business Media LLC, 2024
  • Published in: Nature Communications, 15 (2024) 1
  • Language: English
  • DOI: 10.1038/s41467-023-44339-z
  • ISSN: 2041-1723
  • Keywords: General Physics and Astronomy ; General Biochemistry, Genetics and Molecular Biology ; General Chemistry ; Multidisciplinary
  • Origination:
  • Footnote:
  • Description: <jats:title>Abstract</jats:title><jats:p>To curb viral epidemics and pandemics, antiviral drugs are needed with activity against entire genera or families of viruses. Here, we develop a cell-based multiplex antiviral assay for high-throughput screening against multiple viruses at once, as demonstrated by using three distantly related orthoflaviviruses: dengue, Japanese encephalitis and yellow fever virus. Each virus is tagged with a distinct fluorescent protein, enabling individual monitoring in cell culture through high-content imaging. Specific antisera and small-molecule inhibitors are employed to validate that multiplexing approach yields comparable inhibition profiles to single-virus infection assays. To facilitate downstream analysis, a kernel is developed to deconvolute and reduce the multidimensional quantitative data to three cartesian coordinates. The methodology is applicable to viruses from different families as exemplified by co-infections with chikungunya, parainfluenza and Bunyamwera viruses. The multiplex approach is expected to facilitate the discovery of broader-spectrum antivirals, as shown in a pilot screen of approximately 1200 drug-like small-molecules.</jats:p>
  • Access State: Open Access