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Volume 26, Number 9—September 2020
Research

Isolation, Sequence, Infectivity, and Replication Kinetics of Severe Acute Respiratory Syndrome Coronavirus 2

Arinjay Banerjee, Jalees A. Nasir1, Patrick Budylowski1, Lily Yip, Patryk Aftanas, Natasha Christie, Ayoob Ghalami, Kaushal Baid, Amogelang R. Raphenya, Jeremy A. Hirota, Matthew S. Miller, Allison J. McGeer, Mario Ostrowski, Robert A. Kozak1, Andrew G. McArthur, Karen MossmanComments to Author , and Samira Mubareka
Author affiliations: McMaster University, Hamilton, Ontario, Canada (A. Banerjee, J.A. Nasir, K. Baid, A.R. Raphenya, J.A. Hirota, M.S. Miller, A.G. McArthur, K. Mossman); University of Toronto, Toronto, Ontario, Canada (P. Budylowski, N. Christie, A. Ghalami, A.J. McGeer, M. Ostrowski, R.A. Kozak, S. Mubareka); Sunnybrook Research Institute, Toronto (L. Yip, P. Aftanas, R.A. Kozak, S. Mubareka); Mount Sinai Hospital, Toronto (A.J. McGeer)

Main Article

Figure 2

Isolating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from patients with coronavirus disease (COVID-19). A) Vero E6 cells were mock inoculated or inoculated with midturbinate clinical specimens from COVID-19 patients. Cells were incubated for 72 h and observed for cytopathic effect (CPE) under a light microscope. Original magnification ×10. B) To determine if supernatant from Vero E6 cells that were mock inoculated or inoculated with clinical specimens contained replication comp

Figure 2. Isolating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from patients with coronavirus disease (COVID-19). A) Vero E6 cells were mock inoculated or inoculated with midturbinate clinical specimens from COVID-19 patients. Cells were incubated for 72 h and observed for cytopathic effect (CPE) under a light microscope. Original magnification ×10. B) To determine if supernatant from Vero E6 cells that were mock inoculated or inoculated with clinical specimens contained replication competent virus, we reinoculated a fresh monolayer of Vero E6 cells and observed cells under a light microscope for CPE after 24 h. Original magnification ×10. C) Quantitative real-time PCR was used to detect SARS-CoV-2 5′-UTR and E gene in RNA extracted from supernatant that was collected from Vero E6 cells that were mock infected or infected with clinical specimens from COVID-19 patients for 72 h. D) Electron micrograph of Vero E6 cells that were reinfected for 48 h with supernatant that was collected from Vero E6 cells infected with clinical specimens. Original magnification ×36,000. Inset, zoomed and cropped from the original electron micrograph, shows coronavirus-like particles. M, mock specimen; specimen 1, SARS-CoV-2/SB2; specimen 2, SARS-CoV-2/SB3-TYAGNC. E, envelope; UTR, untranslated region.

Main Article

1These authors contributed equally to this article.

Page created: June 03, 2020
Page updated: June 19, 2020
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