Growth, detection, quantification, and inactivation of SARS-CoV-2

Virology
James Brett CaseMichael S Diamond

Abstract

Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 is the agent responsible for the coronavirus disease 2019 (COVID-19) global pandemic. SARS-CoV-2 is closely related to SARS-CoV, which caused the 2003 SARS outbreak. Although numerous reagents were developed to study SARS-CoV infections, few have been applicable to evaluating SARS-CoV-2 infection and immunity. Current limitations in studying SARS-CoV-2 include few validated assays with fully replication-competent wild-type virus. We have developed protocols to propagate, quantify, and work with infectious SARS-CoV-2. Here, we describe: (1) virus stock generation, (2) RT-qPCR quantification of SARS-CoV-2 RNA; (3) detection of SARS-CoV-2 antigen by flow cytometry, (4) quantification of infectious SARS-CoV-2 by focus-forming and plaque assays; and (5) validated protocols for virus inactivation. Collectively, these methods can be adapted to a variety of experimental designs, which should accelerate our understanding of SARS-CoV-2 biology and the development of effective countermeasures against COVID-19.

References

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Citations

Dec 30, 2020·Transboundary and Emerging Diseases·Yohannes BerhaneBradley Pickering
Oct 14, 2020·The Journal of Infection·Kieran A WalshMichelle O'Neill
Feb 6, 2021·Cell Reports·Timothy A BatesFikadu G Tafesse
Jan 13, 2021·Methods and Protocols·Kaitlyn A BarrowJason S Debley
Mar 15, 2021·Virology·James S TerryBrian J Geiss
Jun 5, 2021·Proceedings of the National Academy of Sciences of the United States of America·Ron SenderRon Milo
May 27, 2021·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Avrill AsplandMichael Solga
Jun 18, 2021·Journal of Medical Virology·Satoshi HiroiTatsuo Shioda

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Datasets Mentioned

BETA
MG772933

Methods Mentioned

BETA
phage display
flow cytometry
Infection
PCR
in vitro transcription
in
restriction digest
flow-cytometry
FACS

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