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

Viruses
Alexander S JurekaChristopher F Basler

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.

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Citations

Dec 9, 2020·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Avrill M AsplandSherry Thornton
Jan 9, 2021·The Journal of General Virology·Heidi AuerswaldJose A Garcia-Rivera
Jan 20, 2021·Revue des maladies respiratoires·S PilletA Vabret
Jan 6, 2021·Transboundary and Emerging Diseases·Glenn A MarshSeshadri S Vasan
Feb 11, 2021·Archives of Dermatological Research·Heba A Abdelkader
Jan 15, 2021·Clinical Microbiology Reviews·Aleksandra SynowiecKrzysztof Pyrc
Jan 27, 2021·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Daniel GeanonAdeeb H Rahman
Mar 2, 2021·Der Pathologe·N RitschelUNKNOWN „DEFEAT PANDEMIcs – Neuropathologische Referenzdiagnostik bei COVID-19“
Mar 7, 2021·Methods and Protocols·Ranjeet KumarSelvakumar Subbian
Apr 4, 2021·Viruses·Emma K LovedayMatthew P Taylor
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May 22, 2021·Environmental Science : Water Research & Technology·Brian M PecsonUNKNOWN SARS-CoV-2 Interlaboratory Consortium
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
Jul 15, 2021·Chemical Society Reviews·Nan WangTetsuro Majima
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Dec 5, 2021·Scientific Reports·Pooya AfaghiKhashayar Ghandi

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Methods Mentioned

BETA
Assay
light microscopy
electron microscopy

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