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.2021 Sep 10;87(19):e0031421.
doi: 10.1128/AEM.00314-21. Epub 2021 Jul 21.

Heat-Treated Virus Inactivation Rate Depends Strongly on Treatment Procedure: Illustration with SARS-CoV-2

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Heat-Treated Virus Inactivation Rate Depends Strongly on Treatment Procedure: Illustration with SARS-CoV-2

Amandine Gamble et al. Appl Environ Microbiol..

Abstract

Decontamination helps limit environmental transmission of infectious agents. It is required for the safe reuse of contaminated medical, laboratory, and personal protective equipment, and for the safe handling of biological samples. Heat treatment is a common decontamination method, notably used for viruses. We show that for liquid specimens (here, solution of SARS-CoV-2 in cell culture medium), the virus inactivation rate under heat treatment at 70°C can vary by almost two orders of magnitude depending on the treatment procedure, from a half-life of 0.86 min (95% credible interval [CI] 0.09, 1.77) in closed vials in a heat block to 37.04 min (95% CI 12.64, 869.82) in uncovered plates in a dry oven. These findings suggest a critical role of evaporation in virus inactivation via dry heat. Placing samples in open or uncovered containers may dramatically reduce the speed and efficacy of heat treatment for virus inactivation. Given these findings, we reviewed the literature on temperature-dependent coronavirus stability and found that specimen container types, along with whether they are closed, covered, or uncovered, are rarely reported in the scientific literature. Heat-treatment procedures must be fully specified when reporting experimental studies to facilitate result interpretation and reproducibility, and must be carefully considered when developing decontamination guidelines.IMPORTANCE Heat is a powerful weapon against most infectious agents. It is widely used for decontamination of medical, laboratory, and personal protective equipment, and for biological samples. There are many methods of heat treatment, and methodological details can affect speed and efficacy of decontamination. We applied four different heat-treatment procedures to liquid specimens containing SARS-CoV-2. Our results show that the container used to store specimens during decontamination can substantially affect inactivation rate; for a given initial level of contamination, decontamination time can vary from a few minutes in closed vials to several hours in uncovered plates. Reviewing the literature, we found that container choices and heat treatment methods are only rarely reported explicitly in methods sections. Our study shows that careful consideration of heat-treatment procedure-in particular the choice of specimen container and whether it is covered-can make results more consistent across studies, improve decontamination practice, and provide insight into the mechanisms of virus inactivation.

Keywords: SARS-CoV-2; coronavirus; decontamination; environmental persistence; environmental stability; heat treatment; temperature.

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Figures

FIG 1
FIG 1
Inactivation of SARS-CoV-2 by heat treatment under different procedures. (A) A solution of SARS-CoV-2 was exposed to 70°C heat. Samples were placed in uncovered or covered 24-well plates, or in closed 2-ml vials before heat treatment using a dry oven or a heat block containing water. (B) Samples were then collected at the indicated time points during heat treatment. Viable virus titer estimated by endpoint titration is shown in TCID50/ml medium on a logarithmic scale. Points show estimated titers for each collected sample; vertical bar shows a 95% credible interval. Time-points with no positive wells for any replicate are plotted as triangles at the approximate single-replicate detection limit of the assay (LOD, denoted by a black dotted line at 100.5 TCID50/ml medium) to indicate that a range of sub-LOD values are plausible. Lines show predicted decay of virus titer over time (10 random draws per data point from the joint posterior distribution of the slope and intercept). Panel A created withBioRender.com.
FIG 2
FIG 2
Half-life of SARS-CoV-2 in a solution exposed to 70°C heat under different procedures. Quantile dotplots (69) of the posterior distribution for half-life of viable virus under each different heat-treatment procedure. Half-lives were calculated from the estimated exponential decay rates of virus titer (Fig. 1B) and plotted on a logarithmic scale. For each distribution, the black dot shows the posterior median estimate and the black line shows the 95% credible interval.
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