Heat inactivation of the Severe Acute Respiratory Syndrome Coronavirus 2

Batéjat, Christophe; Grassin, Quentin; Manuguerra, Jean-Claude; Leclercq, India

doi:10.1101/2020.05.01.067769

Cited by 58 publications

(67 citation statements)

References 12 publications

(8 reference statements)

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“…Up-front heating of freshly collected saliva samples is attractive as a simple method to inactivate the virus without having to open the collection vessel. Indeed, heat treatment is often used to inactivate saliva patient samples, 29,30 thus conferring added biosafety by decreasing the likelihood of viral transmission via sample handling by personnel. Common conditions for SARS-CoV-2 inactivation are heating at 56-60°C for 30-60 min, 30,31 although other temperature and times have been examined.…”

Section: Development Of a Direct Saliva-to-rt-qpcr Process For Detectmentioning

confidence: 99%

“…As temperature and incubation time were increased, substantial improvement in virus detection was observed, with 100% identification of all SARS-CoV-2 genes, in all replicate samples, being detected following a 30 min incubation at 95°C. Importantly, a short heating time (5 minutes) at 95 o C (as has been examined by others 29,32 ) does not allow for sensitive detection; the 30 minute duration is essential, as it is likely that this extended heating inactivates components of saliva that inhibit RT-qPCR. Thus, proper heating of patient samples allows for virus detection without the need for RNA extraction, with the added benefit of inactivating the samples, thus substantially reducing biohazard risks.…”

Section: Development Of a Direct Saliva-to-rt-qpcr Process For Detectmentioning

confidence: 99%

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Saliva-Based Molecular Testing for SARS-CoV-2 that Bypasses RNA Extraction

Rañoa

Holland

Alnaji

et al. 2020

Preprint

123

178

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Convenient, repeatable, large-scale molecular testing for SARS-CoV-2 would be a key weapon to help control the COVID-19 pandemic. Unfortunately, standard SARS-CoV-2 testing protocols are invasive and rely on numerous items that can be subject to supply chain bottlenecks, and as such are not suitable for frequent repeat testing. Specifically, personal protective equipment (PPE), nasopharyngeal (NP) swabs, the associated viral transport media (VTM), and kits for RNA isolation and purification have all been in short supply at various times during the COVID-19 pandemic. Moreover, SARS-CoV-2 is spread through droplets and aerosols transmitted through person-to-person contact, and thus saliva may be a relevant medium for diagnosing SARS-CoV-2 infection status. Here we describe a saliva-based testing method that bypasses the need for RNA isolation/purification. In experiments with inactivated SARS-CoV-2 virus spiked into saliva, this method has a limit of detection of 500-1000 viral particles per mL, rivalling the standard NP swab method, and initial studies also show excellent performance with 100 clinical samples. This saliva-based process is operationally simple, utilizes readily available materials, and can be easily implemented by existing testing sites, thus allowing for high-throughput, rapid, and repeat testing of large populations. Graphical Abstract3 BackgroundThe slow roll-out and inconsistent availability of diagnostic testing for SARS-CoV-2 has hobbled efforts to control the COVID-19 pandemic in many countries. Testing protocols based on the use of nasopharyngeal (NP) swabs as the collection agent, placed in a tube containing viral transport media (VTM), followed by RNA isolation/purification and subsequent analysis by RT-qPCR is currently the most common method ( Figure 1A). 1,2 While some variant of this process has been implemented worldwide, there are multiple challenges with this workflow. Sample collection using NP swabs requires healthcare workers wearing personal protective equipment (PPE) to collect samples, the swabs can be uncomfortable for the patients during collection, and the swabs and the associated VTM have been in short supply at many times and in most locations. In addition, RNA isolation/purification is another significant bottleneck, both in the time and labor required for this process, and in the availability of the equipment and reagents. All of these components also add to the cost of the testing process.There is emerging consensus that widespread, frequently repeated testing is necessary for a safer return to activities that are important for society. Given the data suggesting that SARS-CoV-2 can be spread by pre-symptomatic/asymptomatic carriers, 3-6 localized outbreaks could be dramatically reduced or prevented if individuals shedding SARS-CoV-2 could be readily identified and isolated. For example, imagine a testing bubble placed over a group that desires face-to-face interaction -employees of a company, members of a sports team, extended family networks, etc. If all members of...

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Section: Development Of a Direct Saliva-to-rt-qpcr Process For Detectmentioning

confidence: 99%

Section: Development Of a Direct Saliva-to-rt-qpcr Process For Detectmentioning

confidence: 99%

Saliva-Based Molecular Testing for SARS-CoV-2 that Bypasses RNA Extraction

Rañoa

Holland

Alnaji

et al. 2020

Preprint

123

178

View full text Add to dashboard Cite

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“…For SARS-CoV-2, after 30 min at 56 °C, the magnitude of the virus titre reduction was 3.0 log 10 [ 33 ], ≥4.6 log 10 [ 37 ], ≥5.0 log 10 [ 66 ], ≥5.9 log 10 [ 67 ], 5.0–6.0 log 10 [ 69 ], and 7.0 log 10 [ 70 ]. At 60 °C, there was a 7.0 log 10 reduction after 15 min [ 70 ], and a 5.0–6.0 log 10 reduction (complete inactivation) in clinical samples after 60 min [ 69 ] ( Table 5 ).…”

Section: Disinfectionmentioning

confidence: 99%

“…At 60 °C, there was a 7.0 log 10 reduction after 15 min [ 70 ], and a 5.0–6.0 log 10 reduction (complete inactivation) in clinical samples after 60 min [ 69 ] ( Table 5 ). According to an unpublished study [ 67 ], at 65 °C, SARS-CoV-2 was inactivated (≥5.5 log 10 reduction) in cell culture, nasopharyngeal samples, and serum after 15, 10, and 10 min, respectively ( Table 5 ). The efficacy of heat treatment against SARS-CoV-2 was also evaluated at other temperatures that varied among studies, including 70 °C [ 37 , 39 , 68 ], 92 °C [ 69 ], 95 °C [ 67 ], and 98 °C [ 66 ] ( Table 5 ).…”

Section: Disinfectionmentioning

confidence: 99%

“…Overall, heat treatment at 60 °C for 60 min would lead to SARS-CoV-1 inactivation according to six studies [ 31 , 34 , 35 , 45 , 50 , 65 ] (~3.5–8.0 log 10 reductions in a variety of media), and to SARS-CoV-2 inactivation as per six studies [ 33 , 37 , 66 , 67 , 69 , 70 ] (~4.6–7.0 log 10 reduction) ( Table 5 ). However, one study showed residual SARS-CoV-1 infectivity until 90 min at 65 °C [ 46 ].…”

Section: Disinfectionmentioning

confidence: 99%

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Rapid Review of SARS-CoV-1 and SARS-CoV-2 Viability, Susceptibility to Treatment, and the Disinfection and Reuse of PPE, Particularly Filtering Facepiece Respirators

Derraik

Anderson

Connelly

et al. 2020

IJERPH

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In the COVID-19 pandemic caused by SARS-CoV-2, hospitals are often stretched beyond capacity. There are widespread reports of dwindling supplies of personal protective equipment (PPE), particularly N95-type filtering facepiece respirators (FFRs), which are paramount to protect frontline medical/nursing staff, and to minimize further spread of the virus. We carried out a rapid review to summarize the existing literature on the viability of SARS-CoV-2, the efficacy of key potential disinfection procedures against the virus (specifically ultraviolet light and heat), and the impact of these procedures on FFR performance, material integrity, and/or fit. In light of the recent discovery of SARS-CoV-2 and limited associated research, our review also focused on the closely related SARS-CoV-1. We propose a possible whole-of-PPE disinfection solution for potential reuse that could be rapidly instituted in many health care settings, without significant investments in equipment.

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Sub‐second heat inactivation of coronavirus using a betacoronavirus model

Jiang

Zhang

Wippold

et al. 2021

Biotech & Bioengineering

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Heat treatment denatures viral proteins that comprise the virion, making the virus incapable of infecting a host. Coronavirus (CoV) virions contain single‐stranded RNA genomes with a lipid envelope and four proteins, three of which are associated with the lipid envelope and thus are thought to be easily denatured by heat or surfactant‐type chemicals. Prior studies have shown that a temperature as low as 75°C with a treatment duration of 15 min can effectively inactivate CoV. The degree of CoV heat inactivation greatly depends on the length of heat treatment time and the temperature applied. With the goal of finding whether sub‐second heat exposure of CoV can sufficiently inactivate CoV, we designed and developed a simple fluidic system that can measure sub‐second heat inactivation of CoV. The system is composed of a stainless‐steel capillary immersed in a temperature‐controlled oil bath followed by an ice bath, through which virus solution can flow at various speeds. Flowing virus solution at different speeds, along with temperature control and monitoring system, allows the virus to be exposed to the desired temperature and treatment durations with high accuracy. Using mouse hepatitis virus, a betacoronavirus, as a model CoV system, we identified that 71.8°C for 0.51 s exposure is sufficient to obtain >5 Log 10 reduction in viral titer (starting titer: 5 × 10 7 PFU/ml), and that when exposed to 83.4°C for 1.03 s, the virus was completely inactivated (>6 Log 10 reduction).

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Heat inactivation of the Severe Acute Respiratory Syndrome Coronavirus 2

Cited by 58 publications

References 12 publications

Saliva-Based Molecular Testing for SARS-CoV-2 that Bypasses RNA Extraction

Saliva-Based Molecular Testing for SARS-CoV-2 that Bypasses RNA Extraction

Rapid Review of SARS-CoV-1 and SARS-CoV-2 Viability, Susceptibility to Treatment, and the Disinfection and Reuse of PPE, Particularly Filtering Facepiece Respirators

Sub‐second heat inactivation of coronavirus using a betacoronavirus model

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