Preprint Brief Report Version 1 Preserved in Portico This version is not peer-reviewed

RNA Extraction Alternative Method for SARS-CoV-2 Molecular Diagnosis

Version 1 : Received: 13 May 2021 / Approved: 14 May 2021 / Online: 14 May 2021 (14:46:42 CEST)

How to cite: Bianchini, E.; Rossignolo, F.; Perfranceschi, M.; Cazzin, C.; Silva, R.; Formenti, F.; Perandin, F.; Griffante, C.; Remelli, R.; Padovani, M.; Scarso, S.; Leonardi, M.; Piubelli, C. RNA Extraction Alternative Method for SARS-CoV-2 Molecular Diagnosis. Preprints 2021, 2021050342 (doi: 10.20944/preprints202105.0342.v1). Bianchini, E.; Rossignolo, F.; Perfranceschi, M.; Cazzin, C.; Silva, R.; Formenti, F.; Perandin, F.; Griffante, C.; Remelli, R.; Padovani, M.; Scarso, S.; Leonardi, M.; Piubelli, C. RNA Extraction Alternative Method for SARS-CoV-2 Molecular Diagnosis. Preprints 2021, 2021050342 (doi: 10.20944/preprints202105.0342.v1).

Abstract

Background: the devastating outbreak of COVID-19 poses serious challenges for the diagnostics laboratories, which are often facing global shortage of reagents and equipment. With the aim of increasing the diagnostic throughput for SARS-CoV-2 molecular test, the purpose of this study was to validate an additional RNA extraction method respect to those already recommended by WHO and the US Centers for Disease Control and Prevention (CDC). Methods: a new protocol for RNA extraction from nasopharyngeal swab was set up, adapting the Qiagen RNeasy 96 plate and validated on a set of 100 clinical samples analyzed in parallel by Roche-Magnapure method (already recommended by CDC guidelines). Results: the internal control and target genes analysis showed a good agreement between the two extraction methods indicating that the two methods can be considered equivalent and that the RNeasy-adapted method can be applied for the SARS-CoV-2 diagnostics. The addition of this new extraction method resulted in a throughput increase for SARS-CoV-2 molecular test of about 2000 samples/month during the initial months of the pandemic emergency in which the lack of reagents for the extraction led to an insufficient sample processing throughput of the analysis of the swabs.

Keywords

COVID-19; SARS-CoV-2; nasopharyngeal swab; RNA extraction; RT-PCR

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