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Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E

RT-qPCR remains a key diagnostic methodology for COVID-19/SARS-CoV-2. Typically, nasal or saliva swabs from patients are placed in virus transport media (VTM), RNA is extracted at the pathology laboratory, and viral RNA is measured using RT-qPCR. In this study, we describe the use of TNA-Cifer Reage...

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Autores principales: Pollak, Nina M., Rawle, Daniel J., Yan, Kexin, Buckley, Cameron, Le, Thuy T., Wang, Claire Y. T., Ertl, Nicole G., van Huyssteen, Karla, Crkvencic, Nicole, Hashmi, Misha, Lyons, Russell E., Whiley, David M., Suhrbier, Andreas, Macdonald, Joanne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10590215/
https://www.ncbi.nlm.nih.gov/pubmed/37869655
http://dx.doi.org/10.3389/fmicb.2023.1238542
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author Pollak, Nina M.
Rawle, Daniel J.
Yan, Kexin
Buckley, Cameron
Le, Thuy T.
Wang, Claire Y. T.
Ertl, Nicole G.
van Huyssteen, Karla
Crkvencic, Nicole
Hashmi, Misha
Lyons, Russell E.
Whiley, David M.
Suhrbier, Andreas
Macdonald, Joanne
author_facet Pollak, Nina M.
Rawle, Daniel J.
Yan, Kexin
Buckley, Cameron
Le, Thuy T.
Wang, Claire Y. T.
Ertl, Nicole G.
van Huyssteen, Karla
Crkvencic, Nicole
Hashmi, Misha
Lyons, Russell E.
Whiley, David M.
Suhrbier, Andreas
Macdonald, Joanne
author_sort Pollak, Nina M.
collection PubMed
description RT-qPCR remains a key diagnostic methodology for COVID-19/SARS-CoV-2. Typically, nasal or saliva swabs from patients are placed in virus transport media (VTM), RNA is extracted at the pathology laboratory, and viral RNA is measured using RT-qPCR. In this study, we describe the use of TNA-Cifer Reagent E in a pre-clinical evaluation study to inactivate SARS-CoV-2 as well as prepare samples for RT-qPCR. Adding 1 part TNA-Cifer Reagent E to 5 parts medium containing SARS-CoV-2 for 10 min at room temperature inactivated the virus and permitted RT-qPCR detection. TNA-Cifer Reagent E was compared with established column-based RNA extraction and purification methodology using a panel of human clinical nasal swab samples (n = 61), with TNA-Cifer Reagent E showing high specificity (100%) and sensitivity (97.37%). Mixtures of SARS-CoV-2 virus and TNA-Cifer Reagent E could be stored for 3 days at room temperature or for 2 weeks at 4°C without the loss of RT-qPCR detection sensitivity. The detection sensitivity was preserved when TNA-Cifer Reagent E was used in conjunction with a range of VTM for saliva samples but only PBS (Gibco) and Amies Orange for nasal samples. Thus, TNA-Cifer Reagent E improves safety by rapidly inactivating the virus during sample processing, potentially providing a safe means for molecular SARS-CoV-2 testing outside traditional laboratory settings. The reagent also eliminates the need for column-based and/or automated viral RNA extraction/purification processes, thereby providing cost savings for equipment and reagents, as well as reducing processing and handling times.
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spelling pubmed-105902152023-10-22 Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E Pollak, Nina M. Rawle, Daniel J. Yan, Kexin Buckley, Cameron Le, Thuy T. Wang, Claire Y. T. Ertl, Nicole G. van Huyssteen, Karla Crkvencic, Nicole Hashmi, Misha Lyons, Russell E. Whiley, David M. Suhrbier, Andreas Macdonald, Joanne Front Microbiol Microbiology RT-qPCR remains a key diagnostic methodology for COVID-19/SARS-CoV-2. Typically, nasal or saliva swabs from patients are placed in virus transport media (VTM), RNA is extracted at the pathology laboratory, and viral RNA is measured using RT-qPCR. In this study, we describe the use of TNA-Cifer Reagent E in a pre-clinical evaluation study to inactivate SARS-CoV-2 as well as prepare samples for RT-qPCR. Adding 1 part TNA-Cifer Reagent E to 5 parts medium containing SARS-CoV-2 for 10 min at room temperature inactivated the virus and permitted RT-qPCR detection. TNA-Cifer Reagent E was compared with established column-based RNA extraction and purification methodology using a panel of human clinical nasal swab samples (n = 61), with TNA-Cifer Reagent E showing high specificity (100%) and sensitivity (97.37%). Mixtures of SARS-CoV-2 virus and TNA-Cifer Reagent E could be stored for 3 days at room temperature or for 2 weeks at 4°C without the loss of RT-qPCR detection sensitivity. The detection sensitivity was preserved when TNA-Cifer Reagent E was used in conjunction with a range of VTM for saliva samples but only PBS (Gibco) and Amies Orange for nasal samples. Thus, TNA-Cifer Reagent E improves safety by rapidly inactivating the virus during sample processing, potentially providing a safe means for molecular SARS-CoV-2 testing outside traditional laboratory settings. The reagent also eliminates the need for column-based and/or automated viral RNA extraction/purification processes, thereby providing cost savings for equipment and reagents, as well as reducing processing and handling times. Frontiers Media S.A. 2023-10-06 /pmc/articles/PMC10590215/ /pubmed/37869655 http://dx.doi.org/10.3389/fmicb.2023.1238542 Text en Copyright © 2023 Pollak, Rawle, Yan, Buckley, Le, Wang, Ertl, van Huyssteen, Crkvencic, Hashmi, Lyons, Whiley, Suhrbier and Macdonald. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Pollak, Nina M.
Rawle, Daniel J.
Yan, Kexin
Buckley, Cameron
Le, Thuy T.
Wang, Claire Y. T.
Ertl, Nicole G.
van Huyssteen, Karla
Crkvencic, Nicole
Hashmi, Misha
Lyons, Russell E.
Whiley, David M.
Suhrbier, Andreas
Macdonald, Joanne
Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
title Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
title_full Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
title_fullStr Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
title_full_unstemmed Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
title_short Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
title_sort rapid inactivation and sample preparation for sars-cov-2 pcr-based diagnostics using tna-cifer reagent e
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10590215/
https://www.ncbi.nlm.nih.gov/pubmed/37869655
http://dx.doi.org/10.3389/fmicb.2023.1238542
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