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

This study highlights the importance of local validation of in-house assays before its availability to the population. Experiments to establish the assay analytical specificity can be easily implemented. At this moment, the proposed method [N1 (primary) and RPDP (modified) (confirmatory)] detected the first case of SARS-CoV-2 Brazilian central-west region, demonstrating that the use of the synthetic RT-qPCR target to investigate novel assays diagnostic parameters in automated workflows is a quick, simple, effective way to be prepared for upcoming threats. The use of spiked samples resembling a real clinical specimen exposes the artificial SARS-CoV-2 sequences to the same background of nucleic acids yields that can be found in the routine and similar amplification behavior of a real SARS-CoV-2 is expected.