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

Henrik Dimke | Zendy; Sanne Løkkegaard Larsen | Zendy; Marianne Nielsine Skov | Zendy; Hanne Larsen | Zendy; Gitte Nyvang Hartmeyer | Zendy; Jesper B. Moeller | Zendy

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

Author(s) -

Henrik Dimke,

Sanne Løkkegaard Larsen,

Marianne Nielsine Skov,

Hanne Larsen,

Gitte Nyvang Hartmeyer,

Jesper B. Moeller

Publication year - 2021

Publication title -

plos one

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.99

H-Index - 332

ISSN - 1932-6203

DOI - 10.1371/journal.pone.0247524

Subject(s) - rna extraction , covid-19 , rna , virology , coronavirus , extraction (chemistry) , chromatography , biology , medicine , chemistry , biochemistry , gene , pathology , disease , infectious disease (medical specialty)

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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