Propagation, Inactivation, and Safety Testing of SARS-CoV-2.

Alexander Jureka,Christopher Basler,Jesus Silvas

doi:10.3390/v12060622

Alexander Jureka, Christopher Basler + Show 1 more

Open Access

https://doi.org/10.3390/v12060622

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Journal: Viruses	Publication Date: Jun 6, 2020
Citations: 111	License type: CC BY 4.0

Affiliation: Georgia State University

Abstract

In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. As work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable the safe study of RNA, DNA, and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.

Highlights

The novel coronavirus SARS-CoV-2, the causative agent of Coronavirus disease 2019 (COVID-19), belongs to the betacoronavirus genus, which includes the highly pathogenic SARS CoV and MERS CoV
The recommendation that experiments involving the propagation of SARs-CoV-2 be performed at biosafety level 3 (BSL3) necessitates the development of methods to safely inactivate the virus and validate inactivation methods to allow an array of studies to be performed at lower biocontainment levels [3,4,5,6]
Despite the number of recent reports in which SARs-CoV-2 has been propagated and quantified, there still remains a lack of general information regarding the temporal cytopathology of SARS-CoV-2 in cell culture

Summary

Introduction

The novel coronavirus SARS-CoV-2, the causative agent of Coronavirus disease 2019 (COVID-19), belongs to the betacoronavirus genus, which includes the highly pathogenic SARS CoV and MERS CoV. As of May 11, 2020, SARS-CoV-2 had caused more than four million infections and more than 250,000 deaths worldwide [2] The emergence of this new virus has prompted urgent worldwide efforts to develop diagnostics, vaccines, and antivirals, to define the natural history of human infection; and to better characterize the virus. Despite the rapidly growing body of literature either deposited on preprint servers or in peer-reviewed scientific journals, there remains a lack of information regarding standardized protocols for work with the virus. Among these needs are the means to grow and quantify infectious virus. Examples include the isolation of RNA from virus and virus-infected cells to characterize viral genome sequences, monitor viral gene

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Conclusion