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

Frontiers in Microbiology

Microbiology

Nicole Crkvencic¹  Misha Hashmi¹  Russell E. Lyons²  Karla van Huyssteen²  Joanne Macdonald³  Nina M. Pollak⁴  Nicole G. Ertl⁵  Cameron Buckley⁵  David M. Whiley⁶  Kexin Yan⁷  Thuy T. Le⁷  Daniel J. Rawle⁷  Andreas Suhrbier⁸  Claire Y. T. Wang⁹ 

[1] Bio Molecular Systems, Potts Point, NSW, Australia;BioCifer Pty Ltd., Auchenflower, QLD, Australia;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;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;Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Herston, QLD, Australia;Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Herston, QLD, Australia;Microbiology Department, Pathology Queensland, Herston, QLD, Australia;Inflammation Biology Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia;Inflammation Biology Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia;GVN Center of Excellence, Australian Infectious Disease Research Centre, Herston, QLD, Australia;Queensland Paediatric Infectious Diseases Laboratory, Centre for Children's Health Research, Brisbane, QLD, Australia;

关键词: COVID-19;    SARS-CoV-2;    RT-qPCR;    diagnostics;    virus inactivation;    safety;    RNA extraction;   

DOI  :  10.3389/fmicb.2023.1238542

 received in 2023-06-12, accepted in 2023-09-07,  发布年份 2023

来源: Frontiers

【 摘 要 】

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.

【 授权许可】

Unknown   
Copyright © 2023 Pollak, Rawle, Yan, Buckley, Le, Wang, Ertl, van Huyssteen, Crkvencic, Hashmi, Lyons, Whiley, Suhrbier and Macdonald.