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Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection

The degradation of SARS-CoV-2 specific ribonucleic acid (RNA) was investigated by a numerical modeling approach based on nucleic acid amplification test (NAAT) results utilizing the SmartAmp technique. The precision of the measurement was verified by the relative standard deviation (RSD) of repeated...

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Autores principales: Takeuchi, Katsuyuki, Yanagisawa, Hiroyuki, Kurosawa, Yukiko, Iida, Yoritsugu, Kawai, Kosuke, Fujimaki, Shigehiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8916628/
https://www.ncbi.nlm.nih.gov/pubmed/35275928
http://dx.doi.org/10.1371/journal.pone.0264541
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author Takeuchi, Katsuyuki
Yanagisawa, Hiroyuki
Kurosawa, Yukiko
Iida, Yoritsugu
Kawai, Kosuke
Fujimaki, Shigehiko
author_facet Takeuchi, Katsuyuki
Yanagisawa, Hiroyuki
Kurosawa, Yukiko
Iida, Yoritsugu
Kawai, Kosuke
Fujimaki, Shigehiko
author_sort Takeuchi, Katsuyuki
collection PubMed
description The degradation of SARS-CoV-2 specific ribonucleic acid (RNA) was investigated by a numerical modeling approach based on nucleic acid amplification test (NAAT) results utilizing the SmartAmp technique. The precision of the measurement was verified by the relative standard deviation (RSD) of repeated measurements at each calibration point. The precision and detection limits were found to be 6% RSD (seven repeated measurements) and 94 copies/tube, respectively, at the lowest calibration point. RNA degradation curves obtained from NAAT data on four different temperatures were in good agreement with the first-order reaction model. By referring to rate constants derived from the results, the Arrhenius model was applied to predict RNA degradation behavior. If the initial RNA concentration was high enough, such as in samples taken from infected bodies, the NAAT results were expected to be positive during testing. On the other hand, if initial RNA concentrations were relatively low, such as RNA in residual viruses on environmental surfaces, special attention should be paid to avoid false-negative results. The results obtained in this study provide a practical guide for RNA sample management in the NAAT of non-human samples.
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spelling pubmed-89166282022-03-12 Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection Takeuchi, Katsuyuki Yanagisawa, Hiroyuki Kurosawa, Yukiko Iida, Yoritsugu Kawai, Kosuke Fujimaki, Shigehiko PLoS One Research Article The degradation of SARS-CoV-2 specific ribonucleic acid (RNA) was investigated by a numerical modeling approach based on nucleic acid amplification test (NAAT) results utilizing the SmartAmp technique. The precision of the measurement was verified by the relative standard deviation (RSD) of repeated measurements at each calibration point. The precision and detection limits were found to be 6% RSD (seven repeated measurements) and 94 copies/tube, respectively, at the lowest calibration point. RNA degradation curves obtained from NAAT data on four different temperatures were in good agreement with the first-order reaction model. By referring to rate constants derived from the results, the Arrhenius model was applied to predict RNA degradation behavior. If the initial RNA concentration was high enough, such as in samples taken from infected bodies, the NAAT results were expected to be positive during testing. On the other hand, if initial RNA concentrations were relatively low, such as RNA in residual viruses on environmental surfaces, special attention should be paid to avoid false-negative results. The results obtained in this study provide a practical guide for RNA sample management in the NAAT of non-human samples. Public Library of Science 2022-03-11 /pmc/articles/PMC8916628/ /pubmed/35275928 http://dx.doi.org/10.1371/journal.pone.0264541 Text en © 2022 Takeuchi et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Takeuchi, Katsuyuki
Yanagisawa, Hiroyuki
Kurosawa, Yukiko
Iida, Yoritsugu
Kawai, Kosuke
Fujimaki, Shigehiko
Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection
title Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection
title_full Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection
title_fullStr Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection
title_full_unstemmed Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection
title_short Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection
title_sort degradation of sars-cov-2 specific ribonucleic acid in samples for nucleic acid amplification detection
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8916628/
https://www.ncbi.nlm.nih.gov/pubmed/35275928
http://dx.doi.org/10.1371/journal.pone.0264541
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