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

Autor:	Pollak NM; Center for Bioinnovation, University of the Sunshine Coast, Sippy Downs, QLD, Australia.; School of Science, Technology and Engineering, University of the Sunshine Coast, Sippy Downs, QLD, Australia.; DMTC Limited, Kew, VIC, Australia., Rawle DJ; Inflammation Biology Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia., Yan K; Inflammation Biology Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia., Buckley C; Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Herston, QLD, Australia., Le TT; Inflammation Biology Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia., Wang CYT; Queensland Paediatric Infectious Diseases Laboratory, Centre for Children's Health Research, Brisbane, QLD, Australia., Ertl NG; Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Herston, QLD, Australia., van Huyssteen K; BioCifer Pty Ltd., Auchenflower, QLD, Australia., Crkvencic N; Bio Molecular Systems, Potts Point, NSW, Australia., Hashmi M; Bio Molecular Systems, Potts Point, NSW, Australia., Lyons RE; BioCifer Pty Ltd., Auchenflower, QLD, Australia., Whiley DM; Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Herston, QLD, Australia.; Microbiology Department, Pathology Queensland, Herston, QLD, Australia., Suhrbier A; Inflammation Biology Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia.; GVN Center of Excellence, Australian Infectious Disease Research Centre, Herston, QLD, Australia., Macdonald J; Center for Bioinnovation, University of the Sunshine Coast, Sippy Downs, QLD, Australia.; School of Science, Technology and Engineering, University of the Sunshine Coast, Sippy Downs, QLD, Australia.; BioCifer Pty Ltd., Auchenflower, QLD, Australia.
Jazyk:	angličtina
Zdroj:	Frontiers in microbiology [Front Microbiol] 2023 Oct 06; Vol. 14, pp. 1238542. Date of Electronic Publication: 2023 Oct 06 (Print Publication: 2023).
DOI:	10.3389/fmicb.2023.1238542
Abstrakt:	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. Competing Interests: NP and NE were post-doctoral research scientists funded by DMTC Ltd., Australia. JM is a Project Leader for DMTC Ltd., Australia and co-founder, shareholder, and director of BioCifer Pty. Ltd., who has licensed the technology. DR and AS were paid consultants for BioCifer. QIMR Berghofer MRI staff were funded via R&D contracts from BioCifer. KH and RL are employees of BioCifer. MH and NC are employees of Bio Molecular Systems. Information in this paper has been included in an International Patent Application PCT/AU2022/051506 (PCT filing date 14th Dec 2022; Applicants BioCifer Pty Ltd, University of the Sunshine Coast, DMTC Limited; Title: Sample preparation reagents; Inventors JM, NP, and NE). The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2023 Pollak, Rawle, Yan, Buckley, Le, Wang, Ertl, van Huyssteen, Crkvencic, Hashmi, Lyons, Whiley, Suhrbier and Macdonald.)
Databáze:	MEDLINE
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