tetano
Editor, Senior Moderator
PLoS One
. 2021 Feb 24;16(2):e0247524.
doi: 10.1371/journal.pone.0247524. eCollection 2021.
Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems
Henrik Dimke[SUP] 1 2 [/SUP], Sanne L Larsen[SUP] 3 [/SUP], Marianne N Skov[SUP] 3 [/SUP], Hanne Larsen[SUP] 3 [/SUP], Gitte N Hartmeyer[SUP] 3 [/SUP], Jesper B Moeller[SUP] 4 5 [/SUP]
Affiliations
Abstract
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.
. 2021 Feb 24;16(2):e0247524.
doi: 10.1371/journal.pone.0247524. eCollection 2021.
Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems
Henrik Dimke[SUP] 1 2 [/SUP], Sanne L Larsen[SUP] 3 [/SUP], Marianne N Skov[SUP] 3 [/SUP], Hanne Larsen[SUP] 3 [/SUP], Gitte N Hartmeyer[SUP] 3 [/SUP], Jesper B Moeller[SUP] 4 5 [/SUP]
Affiliations
- PMID: 33626071
- DOI: 10.1371/journal.pone.0247524
Abstract
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.