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

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 pandem...

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Bibliographic Details
Published in:Viruses Vol. 12; no. 6; p. 622
Main Authors: Jureka, Alexander, Silvas, Jesus, Basler, Christopher
Format: Journal Article
Language:English
Published: Switzerland MDPI AG 06.06.2020
MDPI
Subjects:
agarose
Animals
Betacoronavirus - drug effects
Betacoronavirus - growth & development
Betacoronavirus - pathogenicity
Betacoronavirus - physiology
biosafety
Cell lines
Cellulose
China
Chlorocebus aethiops
Coronaviridae
coronavirus
Coronavirus Infections - virology
Coronaviruses
COVID-19
COVID-19 infection
Culture Media - chemistry
DNA
Formaldehyde
formalin
Gene expression
Genomes
Guanidines - pharmacology
Heat
HEK293 Cells
Humans
inactivation
Infections
Infectivity
lactones
pandemic
Pandemics
pathogenicity
Phenols - pharmacology
Plaque assay
Pneumonia, Viral - virology
Propagation
Propiolactone - pharmacology
Ribonucleic acid
RNA
safety testing
SARS-CoV-2
Sepharose
Severe acute respiratory syndrome coronavirus 2
Vero Cells
Viral Plaque Assay - methods
virology
virus
Virus Inactivation
Viruses
United States > US
China
coronavirus
SARS-CoV-2
virology
virus
plaque assay
inactivation
ISSN:1999-4915, 1999-4915
Online Access:Get full text
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Summary: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.
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Those authors contributed equally to this manuscript.
ISSN:1999-4915
1999-4915
DOI:10.3390/v12060622