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

Henrik Dimke; Sanne L. Larsen; Marianne N. Skov; Hanne Larsen; Gitte N. Hartmeyer; Jesper B. Moeller

doi:10.1371/journal.pone.0247524

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

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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.

Originalsprog	Engelsk
Artikelnummer	e0247524
Tidsskrift	PLOS ONE
Vol/bind	16
Udgave nummer	2
Antal sider	6
ISSN	1932-6203
DOI	https://doi.org/10.1371/journal.pone.0247524
Status	Udgivet - 24. feb. 2021

Bibliografisk note

Funding Information:
The authors received no specific funding for this work.

Finansiering

The authors received no specific funding for this work.

Dokumenter og links

10.1371/journal.pone.0247524Licens: CC BY

Open Access VersionForlagets udgivne version, 792 KBLicens: CC BY

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Citationsformater

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

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{\textregistered}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.",

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author = "Henrik Dimke and Larsen, \{Sanne L.\} and Skov, \{Marianne N.\} and Hanne Larsen and Hartmeyer, \{Gitte N.\} and Moeller, \{Jesper B.\}",

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AU - Dimke, Henrik

AU - Larsen, Sanne L.

AU - Skov, Marianne N.

AU - Larsen, Hanne

AU - Hartmeyer, Gitte N.

AU - Moeller, Jesper B.

N1 - Funding Information: The authors received no specific funding for this work.

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AB - 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.

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KW - Real-Time Polymerase Chain Reaction/methods

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KW - Sensitivity and Specificity

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

Abstract

Bibliografisk note

Finansiering

Dokumenter og links

SDU link

Fingeraftryk

Citationsformater