Isolation and rapid sharing of the 2019 novel coronavirus ( SARS ‐CoV‐2) from the first patient diagnosed with COVID ‐19 in Australia

Abstract

Objectives: To describe the first isolation and sequencing of SARS-CoV-2 in Australia and rapid sharing of the isolate.

SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology

“…To verify the identity and genome sequence of the SARS-CoV-2/p2 virus stock, we isolated genomic RNA from culture supernatant and applied next-generation sequencing (NGS; see methods for details). The resulting consensus sequence was found to be identical to the sequence previously deposited in GenBank (accession number MT007544.1) (34), with one exception (see below). Compared to the SARS-CoV-2 GenBank reference sequence (NC_045512.3) (38) and other field isolates (29), isolate BetaCoV/Australia/VIC01/2020 exhibits >99.9% sequence identity.…”

“…SARS-CoV-2 isolate BetaCoV/Australia/VIC01/2020 was received as a stock derived from two consecutive passages in Vero/hSLAM cells (34). The virus was then propagated two more times at low MOI in Vero E6 cells, in which it caused a severe cytopathic effect (CPE).…”

“…Vero E6 cells and HuH7 cells were grown as described previously (35). SARS-CoV-2 isolate Australia/VIC01/2020 (GeneBank ID: MT007544.1; (34)) was derived from a positively-testing nasopharyngeal swab in Melbourne, Australia, and was propagated twice in Vero/hSLAM cells, before being shared with other laboratories. In Leiden, the virus was passaged two more times at low multiplicity of infection (m.o.i.)…”

“…From a biological and evolutionary point of view, this would seem a strangely anthropocentric consideration, but it ultimately forced us to reach out across the globe to Australian colleagues in Melbourne. After checking our credentials and completing a basic material transfer agreement, they provided us (within one week) with their first SARS-CoV-2 isolate (originally named 2019-nCoV/Victoria/1/2020 and subsequently renamed BetaCoV/Australia/VIC01/2020; (34), which will be used throughout this study. Until now, this isolate has been provided to 17 other laboratories worldwide to promote the rapid characterization of SARS-CoV-2, in this critical time of lockdowns and other preventive measures to avoid a collapse of public health systems.…”

Policy opportunities to enhance sharing for pandemic research

“…These genetic sequences have helped to track the spread of SARS-CoV-2, determine which containment strategies have been successful, and monitor the emergence of adaptive mutations in the viral genome (1). Physical samples of SARS-CoV-2 were, however, unavailable until researchers in Australia isolated the virus from a traveler from Wuhan on 29 January and sent the isolate to the WHO and other laboratories (4).…”