The effect of heat-treatment on SARS-CoV-2 viability and detection

•SARS-CoV-2 is not fully inactivated by heating to 56 °C or 60 °C for up to 60 min.•Complete inactivation occurred at 80 °C for 90 min or 95 °C for 1 or 5 min.•Heating SARS-CoV-2 to 80 °C for more than 30 min results in increased Ct values.•Significant variation in inactivation efficacy occurred bet...

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Bibliographic Details
Published in:Journal of virological methods Vol. 290; p. 114087
Main Authors: Burton, Jane, Love, Hannah, Richards, Kevin, Burton, Christopher, Summers, Sian, Pitman, James, Easterbrook, Linda, Davies, Katherine, Spencer, Peter, Killip, Marian, Cane, Patricia, Bruce, Christine, Roberts, Allen D.G.
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 01.04.2021
Published by Elsevier B.V
Subjects:
biosafety
COVID-19 - diagnosis
COVID-19 Nucleic Acid Testing
heat
Heat inactivation
heat treatment
Hot Temperature
Humans
SARS-CoV-2
SARS-CoV-2 - isolation & purification
SARS-CoV-2 - physiology
Sensitivity and Specificity
Severe acute respiratory syndrome coronavirus 2
temperature
Time Factors
viability
viral load
virus culture
Virus Inactivation
viruses
Heat inactivation
SARS-CoV-2
ISSN:0166-0934, 1879-0984, 1879-0984
Online Access:Get full text
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Summary:•SARS-CoV-2 is not fully inactivated by heating to 56 °C or 60 °C for up to 60 min.•Complete inactivation occurred at 80 °C for 90 min or 95 °C for 1 or 5 min.•Heating SARS-CoV-2 to 80 °C for more than 30 min results in increased Ct values.•Significant variation in inactivation efficacy occurred between replicate samples.•Due to variability heat-inactivation protocols require local validation. The development of safe diagnostic protocols for working with SARS-CoV-2 clinical samples at Biosafety Level 2 (BSL2) requires understanding of the effect of heat-treatment on SARS-CoV-2 viability and downstream RT-PCR sensitivity. In this study heating SARS-CoV-2/England/2/2020 to 56 °C and 60 °C for 15, 30 and 60 min reduced the virus titre by between 2.1 and 4.9 log10 pfu/mL (as determined by plaque assay). Complete inactivation did not occur and there was significant variability between replicates. Viable virus was detected by plaque assay after heat-treatment at 80 °C for 15 or 30 min but not 60 or 90 min. After heat-treatment at 80 °C for 60 min infectious virus was only detected by more sensitive virus culture. No viable virus was detected after heating to 80 °C for 90 min or 95 °C for 1 or 5 min. RT-PCR sensitivity was not compromised by heating to 56 °C and 60 °C. However, RT-PCR sensitivity was reduced (≥3 Ct value increase) after heating the virus to 80 °C for 30 min or longer, or 95 °C for 1 or 5 min. In summary we found that the efficacy of heat-inactivation varies greatly depending on temperature and duration. Local validation of heat-inactivation and its effects downstream is therefore essential for molecular testing.
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ISSN:0166-0934
1879-0984
1879-0984
DOI:10.1016/j.jviromet.2021.114087