Effect of heat inactivation and bulk lysis on real-time reverse transcription PCR detection of the SARS-COV-2: an experimental study
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Abstract
Objective
This study aimed to investigate the effect of heat inactivation and chemical bulklysis on SARS-CoV-2 detection.
Results
About 6.2% (5/80) of samples were changed to negative results in heat inactivation at 60 °C and about 8.7% (7/80) of samples were changed to negative in heat inactivation at 100 °C. The Ct values of heat-inactivated samples (at 60 °C, at 100 °C, and bulk lysis) were significantly different from the temperature at 56 °C. The effect of heat on Ct value should be considered when interpreting diagnostic PCR results from clinical samples which could have an initial low virus concentration. The efficacy of heat-inactivation varies greatly depending on temperature and duration. Local validation of heat-inactivation and its effects is therefore essential for molecular testing.
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SciScore for 10.1101/2022.04.04.22273334: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization Positive samples with known Ct value were selected by simple random sampling technique, from 9,520 positive samples within one month, which is obtained from 28.4% prevalence of Covid-19 in Ethiopia as of August 20/2020. Blinding not detected. Power Analysis not detected. Table 2: Resources
Software and Algorithms Sentences Resources After viral heat inactivation, NA extraction was done from 0.6 ml sample volume on the Abbott m2000SP instrument by using the Abbott mSample Preparation System Deoxyribonucleic acid (DNA) according to the manufacturer’s recommendations [1]. Abbottsuggested: (Abbott, RRID:SCR_010477)Results from OddPub: We …
SciScore for 10.1101/2022.04.04.22273334: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization Positive samples with known Ct value were selected by simple random sampling technique, from 9,520 positive samples within one month, which is obtained from 28.4% prevalence of Covid-19 in Ethiopia as of August 20/2020. Blinding not detected. Power Analysis not detected. Table 2: Resources
Software and Algorithms Sentences Resources After viral heat inactivation, NA extraction was done from 0.6 ml sample volume on the Abbott m2000SP instrument by using the Abbott mSample Preparation System Deoxyribonucleic acid (DNA) according to the manufacturer’s recommendations [1]. Abbottsuggested: (Abbott, RRID:SCR_010477)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:Limitation of the study: Performance has only been established with the specimen types listed in the Intended Use. Other specimen types have not been evaluated and should not be used with this assay. The small sample size can affect the conclusion. Since our study conducted analysis from stored positive samples, weak positive samples may be affected by repeated thaw and refreezing.
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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