A nanoluciferase SARS-CoV-2 for rapid neutralization testing and screening of anti-infective drugs for COVID-19

Xuping Xie (), Antonio E. Muruato, Xianwen Zhang, Kumari G. Lokugamage, Camila R. Fontes-Garfias, Jing Zou, Jianying Liu, Ping Ren, Mini Balakrishnan, Tomas Cihlar, Chien-Te K. Tseng, Shinji Makino, Vineet D. Menachery, John P. Bilello () and Pei-Yong Shi ()
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Xuping Xie: University of Texas Medical Branch
Antonio E. Muruato: University of Texas Medical Branch
Xianwen Zhang: University of Texas Medical Branch
Kumari G. Lokugamage: University of Texas Medical Branch
Camila R. Fontes-Garfias: University of Texas Medical Branch
Jing Zou: University of Texas Medical Branch
Jianying Liu: University of Texas Medical Branch
Ping Ren: University of Texas Medical Branch
Mini Balakrishnan: Gilead Sciences, Inc.
Tomas Cihlar: Gilead Sciences, Inc.
Chien-Te K. Tseng: University of Texas Medical Branch
Shinji Makino: University of Texas Medical Branch
Vineet D. Menachery: University of Texas Medical Branch
John P. Bilello: Gilead Sciences, Inc.
Pei-Yong Shi: University of Texas Medical Branch

Nature Communications, 2020, vol. 11, issue 1, 1-11

Abstract: Abstract A high-throughput platform would greatly facilitate coronavirus disease 2019 (COVID-19) serological testing and antiviral screening. Here we present a high-throughput nanoluciferase severe respiratory syndrome coronavirus 2 (SARS-CoV-2-Nluc) that is genetically stable and replicates similarly to the wild-type virus in cell culture. SARS-CoV-2-Nluc can be used to measure neutralizing antibody activity in patient sera within 5 hours, and it produces results in concordance with a plaque reduction neutralization test (PRNT). Additionally, using SARS-CoV-2-Nluc infection of A549 cells expressing human ACE2 receptor (A549-hACE2), we show that the assay can be used for antiviral screening. Using the optimized SARS-CoV-2-Nluc assay, we evaluate a panel of antivirals and other anti-infective drugs, and we identify nelfinavir, rupintrivir, and cobicistat as the most selective inhibitors of SARS-CoV-2-Nluc (EC50 0.77 to 2.74 µM). In contrast, most of the clinically approved antivirals, including tenofovir alafenamide, emtricitabine, sofosbuvir, ledipasvir, and velpatasvir were inactive at concentrations up to 10 µM. Collectively, this high-throughput platform represents a reliable tool for rapid neutralization testing and antiviral screening for SARS-CoV-2.

Date: 2020
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DOI: 10.1038/s41467-020-19055-7

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