Sub‐second heat inactivation of coronavirus using a betacoronavirus model

Jiang, Yuqian; Zhang, Han; Wippold, Jose A; Gupta, Jyotsana; Dai, Jing; Figueiredo, Paul de; Leibowitz, Julian L.; Han, Arum

doi:10.1002/bit.27720

Cited by 12 publications

(6 citation statements)

References 36 publications

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“…We have created a flowchart of the different processing steps and combinations in our pipeline (Figure 1), which we subsequently explain in more detail. We employed heat inactivation vs non heat inactivation 24 ; compared three different RNA extraction kits (blue) followed by three RT-qPCR mixes and three sets of primers (green).…”

Section: Resultsmentioning

confidence: 99%

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Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation

Lista

Matos

Maguire

et al. 2020

Preprint

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There is a worldwide shortage of reagents to perform detection of SARS-CoV-2. Many clinical diagnostic laboratories rely on commercial platforms that provide integrated end-to-end solutions. While this provides established robust pipelines, there is a clear bottleneck in the supply of reagents given the current situation of extraordinary high demand. Some laboratories resort to implementing kit-free handling procedures, but many other small laboratories will not have the capacity to develop those and/or will perform manual handling of their samples. In order to provide multiple workflows for SARS-CoV-2 nucleic acid detection we compared several commercially available RNA extraction methods: QIAamp Viral RNA Mini Kit (QIAGEN), the recently developed RNAdvance Blood (Beckman) and Mag-Bind Viral DNA/RNA 96 Kit (Omega Bio-tek). We also compared different 1-step RT-qPCR Master Mix brands: TaqMan™ Fast Virus 1-Step Master Mix (ThermoFisher Scientific), qPCRBIO Probe 1-Step Go Lo-ROX (PCR Biosystems) and Luna® Universal Probe One-Step RT-qPCR Kit (NEB). We used the Centre for Disease Control (CDC) recommended primers that detect two regions of the viral N gene as well as those that detect the RdRP gene region as per Public Health England (PHE) guidelines (Charité/WHO/PHE). Our data show that the RNA extraction methods provide similar results. Amongst the qPCR reagents tested, TaqMan™ Fast Virus 1-Step Master Mix and Luna® Universal Probe One-Step RT-qPCR Kit proved most sensitive. The N1 and N2 primer-probes provide a more reliable detection than the RdRP_SARSr primer-probe set, particularly in samples with low viral titres. Importantly, we have implemented a protocol using heat inactivation and demonstrate that it has minimal impact on the sensitivity of the qPCR in clinical samples -potentially making SARS-CoV-2 testing portable to settings that do not have CL-3 facilities. In summary, we provide several testing pipelines that can be easily implemented in other laboratories and have made all our protocols and SOPs freely available at https://osf.io/uebvj/.

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Section: Resultsmentioning

confidence: 99%

“…Representation of our workflow. We employed heat inactivation vs non heat inactivation24 ; compared three different RNA extraction kits (blue) followed by three RT-qPCR mixes and three sets of primers (green).…”

mentioning

confidence: 99%

Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation

Lista

Matos

Maguire

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…Rather, heating to 80 °C for 90 min or 95 °C for 1 or 5 min can obtain full inactivation along with minimized RT-PCR sensitivity through increased cycle threshold (ct) values, indicating lowered viral load ( Burton et al., 2021 ). Moreover, the heat inactivation of coronaviruses at the sub-second level, namely the complete inactivation of the Murine coronavirus (MHC) model at 83.4 °C in 1.03 s potentially consolidates the theorized effect of steam and heat against SARS-CoV-2 ( Jiang et al., 2021 ).…”

Section: Introductionmentioning

confidence: 92%

Theoretical effectiveness of steam inhalation against SARS-CoV-2 infection: updates on clinical trials, mechanism of actions, and traditional approaches

Chowdhury

Alif

Alam

et al. 2022

Heliyon

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Steam inhalation therapy can be a contemporary approach for COVID-19 affected patients of all age groups to manage respiratory conditions, though it presently lacks the scientific backing to establish itself as a befitting practice. The age of COVID-19 has facilitated this traditional home remedy to resurface among the general mass as a helpful approach for the prevention and adjuvant treatment of the disease. In this review, the means of SARS-CoV-2 infection and impact of the parameters, namely steam inhalation and heat on such infection has been delineated via enumerating the effect of the parameters in the human body and against SARS-CoV-2. The literature search was conducted using PubMed, Web of Science, Scopus, ScienceDirect, Wiley Online Library, Google Scholar, and CNKI Scholar databases. The keywords used in the survey include ‘Steam inhalation’, ‘SARS-CoV-2’, ‘COVID-19’, ‘Clinical study’, ‘Mechanism of action’, ‘Traditional uses’, ‘Phytochemistry’ and ‘Adverse effects’. Clinical studies concerning steam inhalation by COVID-19 patients have been comprehended to demarcate the scientific obscurity of the practice. The safety profile of the procedure has also been outlined emphasizing evading measures against COVID-19 and other related disease states. To recapitulate, application of the steam inhalation with herbal concoctions and phytochemicals having folkloric prevalence as an inhalable remedy against respiratory illnesses has been explored in this review work to focus on a new aspect in the COVID-19 treatment paradigm using steam and progress of further research hither.

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“…This was the case Fig 1 . Representation of our workflow. We employed heat inactivation vs non heat inactivation [17]; compared three different RNA extraction kits (blue) followed by three RT-qPCR mixes and three sets of primers (green).…”

Section: Comparison Of Rna Extraction Kitsmentioning

confidence: 99%

Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation

et al. 2021

View full text Add to dashboard Cite

There is a worldwide need for reagents to perform SARS-CoV-2 detection. Some laboratories have implemented kit-free protocols, but many others do not have the capacity to develop these and/or perform manual processing. We provide multiple workflows for SARS-CoV-2 nucleic acid detection in clinical samples by comparing several commercially available RNA extraction methods: QIAamp Viral RNA Mini Kit (QIAgen), RNAdvance Blood/Viral (Beckman) and Mag-Bind Viral DNA/RNA 96 Kit (Omega Bio-tek). We also compared One-step RT-qPCR reagents: TaqMan Fast Virus 1-Step Master Mix (FastVirus, ThermoFisher Scientific), qPCRBIO Probe 1-Step Go Lo-ROX (PCR Biosystems) and Luna® Universal Probe One-Step RT-qPCR Kit (Luna, NEB). We used primer-probes that detect viral N (EUA CDC) and RdRP. RNA extraction methods provided similar results, with Beckman performing better with our primer-probe combinations. Luna proved most sensitive although overall the three reagents did not show significant differences. N detection was more reliable than that of RdRP, particularly in samples with low viral titres. Importantly, we demonstrated that heat treatment of nasopharyngeal swabs at 70°C for 10 or 30 min, or 90°C for 10 or 30 min (both original variant and B 1.1.7) inactivated SARS-CoV-2 employing plaque assays, and had minimal impact on the sensitivity of the qPCR in clinical samples. These findings make SARS-CoV-2 testing portable in settings that do not have CL-3 facilities. In summary, we provide several testing pipelines that can be easily implemented in other laboratories and have made all our protocols and SOPs freely available at https://osf.io/uebvj/.

show abstract

Sub‐second heat inactivation of coronavirus using a betacoronavirus model

Cited by 12 publications

References 36 publications

Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation

Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation

Theoretical effectiveness of steam inhalation against SARS-CoV-2 infection: updates on clinical trials, mechanism of actions, and traditional approaches

Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation

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