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Published VersionCC BY V4.0, Open Access
Journal article
Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
Frontiers in Microbiology, Vol.14, pp.1-12
2023
Appears in COVID-19 Research
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UniSC NewsAbstract
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.
Details
- Title
- Rapid inactivation and sample preparation for SARS-CoV-2 PCR-based diagnostics using TNA-Cifer Reagent E
- Authors
- Nina M. Pollak (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationDaniel J. Rawle (Author) - QIMR Berghofer Medical Research InstituteKexin Yan (Author) - QIMR Berghofer Medical Research InstituteCameron Buckley (Author) - The University of QueenslandThuy T. Le (Author) - QIMR Berghofer Medical Research InstituteClaire Y. T. Wang (Author) - Children’s Health Queensland Hospital and Health ServiceNicole G. Ertl (Author) - The University of QueenslandKarla van Huyssteen (Author) - BioCifer Pty LtdNicole Crkvencic (Author) - Bio Molecular SystemsMisha Hashmi (Author) - Bio Molecular SystemsRussell E. Lyons (Author) - BioCifer Pty LtdDavid M. Whiley (Author) - The University of QueenslandAndreas Suhrbier (Author) - QIMR Berghofer Medical Research InstituteJoanne Macdonald (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
- Publication details
- Frontiers in Microbiology, Vol.14, pp.1-12
- Publisher
- Frontiers Research Foundation
- Date published
- 2023
- DOI
- 10.3389/fmicb.2023.1238542
- ISSN
- 1664-302X
- Copyright note
- © 2023 Pollak, Rawle, Yan, Buckley, Le, Wang, Ertl, van Huyssteen, Crkvencic, Hashmi, Lyons, Whiley, Suhrbier and Macdonald. 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.
- Data Availability
- The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2023.1238542/full#supplementary-material
- Grant note
- DMTC Limited (Australia), the Medical Countermeasures Program (Project 10.75); Bill and Melinda Gates Foundation (OPP1140133); National Health and Medical Research Council (NHMRC) of Australia (APP1173880).
- Organisation Unit
- School of Health - Biomedicine; School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 99971192102621
- Output Type
- Journal article
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