Development of a High-Throughput Sars-Cov-2 Antiviral Testing Method Using Plate-Based Image Cytometry

21 Pages Posted: 23 Mar 2022

See all articles by Laura A. st Clair

Laura A. st Clair

Colorado State University, Fort Collins - Center for Vector-Borne and Infectious Diseases

Leo Li-Ying Chan

affiliation not provided to SSRN

Adam Boretsky

affiliation not provided to SSRN

Bo Lin

affiliation not provided to SSRN

Michael Spedding

affiliation not provided to SSRN

Rushika Perera

Colorado State University, Fort Collins - Center for Vector-Borne and Infectious Diseases

Abstract

The COVID-19 pandemic has created a worldwide public health crisis that has since resulted in 5.8 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) or RT-PCR analysis, but both assays can be tedious and time-consuming, requiring days to visualize and count plaques, or to prepare cell extractions for 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. The Celigo Image Cytometer can provide a more 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.

Note:
Funding Information: This work was funded by The Boettcher Foundation COVID-19 Biomedical Research Innovation Grant to RP, and COVID-19 start-up funds provided to RP by the Department of Microbiology, Immunology and Pathology, and the Office of the Vice President for Research, Colorado State University.

Conflict of Interests: The authors LLC, AB, and BL are employees of Nexcelom Bioscience and declare competing financial interests. The high-throughput image cytometry detection method developed in this manuscript utilizes the Celigo Image Cytometer system from Nexcelom Bioscience. The authors LAS, MS, and RP are named co-inventors on a patent pending covering the novel use of Ambroxol HCl for the treatment of COVID-19 (Demand International n°PCT/EP2021/006314).

Keywords: SARS-CoV-2, COVID-19, antiviral, drug repurposing, image cytometry, Celigo

Suggested Citation

st Clair, Laura A. and Chan, Leo Li-Ying and Boretsky, Adam and Lin, Bo and Spedding, Michael and Perera, Rushika, Development of a High-Throughput Sars-Cov-2 Antiviral Testing Method Using Plate-Based Image Cytometry. Available at SSRN: https://ssrn.com/abstract=4058051 or http://dx.doi.org/10.2139/ssrn.4058051

Laura A. St Clair

Colorado State University, Fort Collins - Center for Vector-Borne and Infectious Diseases ( email )

Department of Economics
Fort Collins, CO 80253-1771
United States

Leo Li-Ying Chan (Contact Author)

affiliation not provided to SSRN ( email )

No Address Available

Adam Boretsky

affiliation not provided to SSRN ( email )

No Address Available

Bo Lin

affiliation not provided to SSRN ( email )

No Address Available

Michael Spedding

affiliation not provided to SSRN

Rushika Perera

Colorado State University, Fort Collins - Center for Vector-Borne and Infectious Diseases ( email )

Department of Economics
Fort Collins, CO 80253-1771
United States

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