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bioRxivMolecular BiologyPreprint

SARS-CoV2 quantification using RT-dPCR: a faster and safer alternative to assist viral genomic copies assessment using RT-qPCR

Paula Rodrigues de AlmeidaiD, Meriane DemolineriD, Ana Karolina Antunes EiseniD, Fágner Henrique HeldtiD, Alana Witt HanseniD, Karoline SchallenbergeriD, Juliane Deise FleckiD, Fernando Rosado SpilkiiD

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Abstract

In this study, serial dilutions of SARS-CoV 2 RNA extract were tested using RT-dPCR using three different primer-probe assays aiming SARS-CoV 2 nucleocapsid coding region. Narrower confidence intervals, indicating high quantification precision were obtained in 100 and 1000-fold serial dilution and RT-dPCR results were equivalent between different assays in the same dilution. High accuracy of this test allowed conclusions regarding the ability of this technique to evaluate precisely the amount of genomic copies present in a sample. We believe that this fast and safe method can assist other researchers in titration of SARS-CoV2 controls used in RT-qPCR without the need of virus isolation.

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