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. 2020 Dec:290:198173.
doi: 10.1016/j.virusres.2020.198173. Epub 2020 Sep 24.

"Sample pooling of RNA extracts to speed up SARS-CoV-2 diagnosis using CDC FDA EUA RT-qPCR kit"

Byron Freire-Paspuel  1 , Patricio Vega-Mariño  2 , Alberto Velez  2 , Marilyn Cruz  2 , Miguel Angel Garcia-Bereguiain  3
Affiliations

"Sample pooling of RNA extracts to speed up SARS-CoV-2 diagnosis using CDC FDA EUA RT-qPCR kit"

Byron Freire-Paspuel et al. Virus Res. 2020 Dec.

Abstract

Background: The CDC protocol for SARS-CoV2 RT-PCR diagnosis (2019-nCoV CDC kit) is considered a gold standard worldwide; based on three different FAM probes (N1 and N2 for viral detection; RP for RNA extraction quality control), three reactions per sample are needed for SARS-CoV-2 diagnosis.

Results: We herein describe a sample pooling protocol: pooling 3 RNA extractions into a single PCR reaction; we tested this protocol with 114 specimens grouped in 38 pools and found no significant differences for N1 and N2 Ct values between pool and single sample PCR reaction.

Conclusion: This pool of three protocol has a sensitivity of 100 % compared to the standard single sample protocol. For a typical 96-well plate, this pool assay allows 96 samples processing, speeding up diagnosis and reducing cost while maintaining clinical performance, particularly useful for SARS-CoV-2 diagnosis at developing countries.

Keywords: CDC; Pools; RT-qPCR; SARS-CoV-2.

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Conflict of interest statement

The authors report no declarations of interest.

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