Analytical sensibility and specificity of two RT-qPCR protocols for SARS-CoV-2 detection performed in an automated workflow

Gustavo Barcelos Barra; Ticiane Henriques Santa Rita; Pedro Goes Mesquita; Rafael Henriques Jacomo; Lidia Freire Abdalla Nery

The RT-qPCR workflow was executed on Flow Flex Solution, which is composed by a liquid handlers and a nucleic acid extractor. The liquid handler pipettes the primary samples and distribute the PCR reaction master mix. Nucleic acids were extracted from 200 ul of the primary sample using MagNA pure 96 DNA and Viral NA Small Volume Kit on Magna pure 96 instrument. One-step RT-qPCR reaction was prepared with LightCycler® Multiplex RNA Virus Master (all from Roche Diagnostics, Mannheim, Germany). Primers/probe concentrations suggested by the interim RT-qPCR protocols providers were maintained and other concentrations were also tested (table 1). The final reaction volume was 10 ul and the automated workflow was able to perform 4 viral assays in 96 samples in 4-5 hours.