Culture of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; f.2019-nCoV) v1

Pyke, Alyssa T.; Mackay, Ian M.; Moore, Frederick; Hurk, Andrew van den; Northill, Judy A; Finger, Mitchell; Simpson, Natalie; Nair, Neelima; Burtonclay, Peter; Moore, Peter F.; Wheatley, Sarah; Moody, Sean; Hall‐Mendelin, Sonja; Gamez, Elisabeth; Jong, Amanda De; Huang, Ben; Taylor, Carmel; Warrilow, David; Genge, Doris; Hewitson, Glen; Sultana, Inga; McMahon, Jamie; Barcelon, Jean

doi:10.17504/protocols.io.bcduis6w

Cited by 4 publications

(6 citation statements)

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“…SARS-CoV-2 stock (strain QLD02/2020, GISAID accession number EPI_ISL_407896) was propagated in Vero E6 cells (ATCC, C1008, CRL-1586, Manassas, USA) as previously described. 6 …”

Section: Methodsmentioning

confidence: 99%

“…SARS-CoV-2 stock (strain QLD02/2020, GISAID accession number EPI_ISL_407896) was propagated in Vero E6 cells (ATCC, C1008, CRL-1586, Manassas, USA) as previously described. 6 Smooth control disks, nanostructured etched Al alloys, and polystyrene tissue culture plate (TCP) surfaces were exposed to 10 μL of viral inoculum (1 × 10 5 TCID 50 /mL) and incubated at room temperature (BSC-2) for the time points 1, 3, 6, 24, and 48 h. Discs were exposed in three independent experiments. To recover live virus, at each respective time point, the exposed surfaces were washed twice by gently pipetting up and down with 32 μL of Opti-MEM reduced serum growth medium (Thermofisher Scientific, Life technologies, Australia Pty Ltd.) 10 times (total volume 64 μL).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

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Antiviral Nanostructured Surfaces Reduce the Viability of SARS-CoV-2

Hasan

Pyke

Nair

et al. 2020

ACS Biomater. Sci. Eng.

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In this letter, we report the ability of the nanostructured aluminum Al 6063 alloy surfaces to inactivate the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). There was no recoverable viable virus after 6 h of exposure to the nanostructured surface, elucidating a 5-log reduction compared to a flat Al 6063 surface. The nanostructured surfaces were fabricated using wet-etching techniques which generated nanotextured, randomly aligned ridges approximately 23 nm wide on the Al 6063 alloy surfaces. In addition to the excellent mechanical resilience properties previously shown, the etched surfaces have also demonstrated superior corrosion resistance compared to the control surfaces. Such nanostructured surfaces have the potential to be used in healthcare environment such as hospitals and public spaces to reduce the surface transmission of SARS-CoV-2 and combat COVID-19.

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“…SARS-CoV-2 stock (strain QLD02/2020, GISAID accession number EPI_ISL_407896) was propagated in Vero E6 cells (ATCC, C1008, CRL-1586, Manassas, USA) as previously described. 6 …”

Section: Methodsmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

Antiviral Nanostructured Surfaces Reduce the Viability of SARS-CoV-2

Hasan

Pyke

Nair

et al. 2020

ACS Biomater. Sci. Eng.

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“…Patient nasopharyngeal swab samples were inoculated onto Vero E6 cell (ATCC, C1008, CRL-1586, Manassas, VA, USA) monolayers as previously described [10]. Passage 2 SARS-CoV-2 stocks were used for the subsequent replication curve kinetic experiments.…”

Section: Patient Samples Cell Culture and Virus Isolationmentioning

confidence: 99%

Replication Kinetics of B.1.351 and B.1.1.7 SARS-CoV-2 Variants of Concern Including Assessment of a B.1.1.7 Mutant Carrying a Defective ORF7a Gene

Pyke

Nair

Hurk

et al. 2021

Viruses

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of COVID-19, is a readily transmissible and potentially deadly pathogen which is currently re-defining human susceptibility to pandemic viruses in the modern world. The recent emergence of several genetically distinct descendants known as variants of concern (VOCs) is further challenging public health disease management, due to increased rates of virus transmission and potential constraints on vaccine effectiveness. We report the isolation of SARS-CoV-2 VOCs imported into Australia belonging to the B.1.351 lineage, first described in the Republic of South Africa (RSA), and the B.1.1.7 lineage originally reported in the United Kingdom, and directly compare the replication kinetics of these two VOCs in Vero E6 cells. In this analysis, we also investigated a B.1.1.7 VOC (QLD1516/2021) carrying a 7-nucleotide deletion in the open reading frame 7a (ORF7a) gene, likely truncating and rendering the ORF7a protein of this virus defective. We demonstrate that the replication of the B.1.351 VOC (QLD1520/2020) in Vero E6 cells can be detected earlier than the B.1.1.7 VOCs (QLD1516/2021 and QLD1517/2021), before peaking at 48 h post infection (p.i.), with significantly higher levels of virus progeny. Whilst replication of the ORF7a defective isolate QLD1516/2021 was delayed longer than the other viruses, slightly more viral progeny was produced by the mutant compared to the unmutated isolate QLD1517/2021 at 72 h p.i. Collectively, these findings contribute to our understanding of SARS-CoV-2 replication and evolutionary dynamics, which have important implications in the development of future vaccination, antiviral therapies, and epidemiological control strategies for COVID-19.

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“…Surfaces have previously been characterized using nanoindentation for mechanical properties, X-ray diffraction for chemical characterization, and contact angle for measuring wettability. 27 32 in DMEM, supplemented with 1 μg/mL tosyl phenylalanyl chloromethyl ketone (TPCK)-treated trypsin (Worthington Biochemical). HCoV-NL63 (Amsterdam-1 strain, provided by Lia van den Hoek, University of Amsterdam) was propagated in LLC-MK2 cells as previously described.…”

Section: Methodsmentioning

confidence: 99%

TiO₂ Nanostructures That Reduce the Infectivity of Human Respiratory Viruses Including SARS-CoV-2

Jaggessar

Velic

Yarlagadda

et al. 2022

ACS Biomater. Sci. Eng.

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The rapid emergence and global spread of the COVID-19 causing Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) and its subsequent mutated strains has caused unprecedented health, economic, and social devastation. Respiratory viruses such as SARS-CoV-2 can be transmitted through both direct and indirect channels, including aerosol respiratory droplets, contamination of inanimate surfaces (fomites), and direct person-to-person contact. Current methods of virus inactivation on surfaces include chemicals and biocides, and while effective, continuous and repetitive cleaning of all surfaces is not always viable. Recent work in the field of biomaterials engineering has established the antibacterial effects of hydrothermally synthesized TiO 2 nanostructured surfaces against both Gram-negative and -positive bacteria. The current study investigates the effectiveness of said TiO 2 nanostructured surfaces against two enveloped human coronaviruses, SARS-CoV-2 and HCoV-NL63, and nonenveloped HRV-16 for surface-based inactivation. Results show that structured surfaces reduced infectious viral loads of SARS-CoV-2 (5 log), HCoV-NL63 (3 log), and HRV-16 (4 log) after 5 h, compared to nonstructured and tissue culture plastic control surfaces. Interestingly, infectious virus remained present on control tissue culture plastic after 7 h exposure. These encouraging results establish the potential use of nanostructured surfaces to reduce the transmission and spread of both enveloped and nonenveloped respiratory viruses, by reducing their infectious period on a surface. The dual antiviral and antibacterial properties of these surfaces support their potential application in a wide variety of settings such as hospitals and healthcare environments, public transport and community hubs.

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Culture of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; f.2019-nCoV) v1

Cited by 4 publications

References 0 publications

Antiviral Nanostructured Surfaces Reduce the Viability of SARS-CoV-2

Antiviral Nanostructured Surfaces Reduce the Viability of SARS-CoV-2

Replication Kinetics of B.1.351 and B.1.1.7 SARS-CoV-2 Variants of Concern Including Assessment of a B.1.1.7 Mutant Carrying a Defective ORF7a Gene

TiO₂ Nanostructures That Reduce the Infectivity of Human Respiratory Viruses Including SARS-CoV-2

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Culture of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; f.2019-nCoV) v1

Cited by 4 publications

References 0 publications

Antiviral Nanostructured Surfaces Reduce the Viability of SARS-CoV-2

Antiviral Nanostructured Surfaces Reduce the Viability of SARS-CoV-2

Replication Kinetics of B.1.351 and B.1.1.7 SARS-CoV-2 Variants of Concern Including Assessment of a B.1.1.7 Mutant Carrying a Defective ORF7a Gene

TiO2 Nanostructures That Reduce the Infectivity of Human Respiratory Viruses Including SARS-CoV-2

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TiO₂ Nanostructures That Reduce the Infectivity of Human Respiratory Viruses Including SARS-CoV-2