Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems

• Main Authors: Dimke, H. (Author), Hartmeyer, G.N (Author), Larsen, H. (Author), Larsen, S.L (Author), Moeller, J.B (Author), Skov, M.N (Author)
• Format: Article
• Language: English
• Published: Public Library of Science 2021
• Subjects: article; chemistry; chloroform; Chloroform; Clinical Laboratory Techniques; COVID-19; COVID-19 Testing; diagnosis; genetics; human; Humans; isolation and purification; laboratory technique; molecular diagnosis; Molecular Diagnostic Techniques; nonhuman; pandemic; Pandemics; phenol; Phenol; procedures; real time polymerase chain reaction; Real-Time Polymerase Chain Reaction; RNA; RNA extraction; RNA purification; RNA, Viral; SARS-CoV-2; sensitivity and specificity; Sensitivity and Specificity; Severe acute respiratory syndrome coronavirus 2; specimen handling; Specimen Handling; swabbing; virology; virus RNA
• Online Access: View Fulltext in Publisher
• ISBN: 19326203 (ISSN)
• DOI: 10.1371/journal.pone.0247524

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use of conventional RNA purification as an alternative to automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical swab specimens were extracted by conventional phenol-chloroform RNA purification and compared to commercial platforms for RNA extraction and the fully integrated Cobas®6800 diagnostic system. Our results show that the conventional RNA extraction is fully comparable to modern automated systems regarding analytical sensitivity and specificity with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, the method is easily scalable and implemented in conventional laboratories as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2. Copyright: © 2021 Dimke et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.