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

Author

Listed:
  • Xuping Xie

    (University of Texas Medical Branch)

  • Antonio E. Muruato

    (University of Texas Medical Branch
    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
    University of Texas Medical Branch
    University of Texas Medical Branch)

  • John P. Bilello

    (Gilead Sciences, Inc.)

  • Pei-Yong Shi

    (University of Texas Medical Branch
    University of Texas Medical Branch
    University of Texas Medical Branch
    University of Texas Medical Branch)

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.

Suggested Citation

  • 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 M, 2020. "A nanoluciferase SARS-CoV-2 for rapid neutralization testing and screening of anti-infective drugs for COVID-19," Nature Communications, Nature, vol. 11(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-19055-7
    DOI: 10.1038/s41467-020-19055-7
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    Cited by:

    1. Jiakai Hou & Yanjun Wei & Jing Zou & Roshni Jaffery & Long Sun & Shaoheng Liang & Ningbo Zheng & Ashley M. Guerrero & Nicholas A. Egan & Ritu Bohat & Si Chen & Caishang Zheng & Xiaobo Mao & S. Stephen, 2024. "Integrated multi-omics analyses identify anti-viral host factors and pathways controlling SARS-CoV-2 infection," Nature Communications, Nature, vol. 15(1), pages 1-14, December.
    2. Yang Liu & Xianwen Zhang & Jianying Liu & Hongjie Xia & Jing Zou & Antonio E. Muruato & Sivakumar Periasamy & Chaitanya Kurhade & Jessica A. Plante & Nathen E. Bopp & Birte Kalveram & Alexander Bukrey, 2022. "A live-attenuated SARS-CoV-2 vaccine candidate with accessory protein deletions," Nature Communications, Nature, vol. 13(1), pages 1-14, December.

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