Litcius/Paper detail

Comparison of Reverse Transcription (RT)-Quantitative PCR and RT-Droplet Digital PCR for Detection of Genomic and Subgenomic SARS-CoV-2 RNA

Sara Morón‐López, Eva Riveira‐Muñoz, Víctor Urrea, Lucía Gutiérrez-Chamorro, Carlos Ávila‐Nieto, Marc Noguera-Julián, Jorge Carrillo, Oriol Mitjà, Lourdes Mateu, Marta Massanella, Ester Ballana, Javier Martínez‐Picado

2023Microbiology Spectrum12 citationsDOIOpen Access PDF

Abstract

We developed one-step reverse transcription (RT)-droplet digital PCR (ddPCR) protocols to detect SARS-CoV-2 RNA and compared them to the gold-standard RT-quantitative PCR (RT-qPCR) method. RT-ddPCR was more sensitive than RT-qPCR in the low-viral-load range, while both techniques were equivalent in the mid- and high-viral-load ranges. Overall, these results suggest that RT-ddPCR might be a viable alternative to RT-qPCR when it comes to detecting low viral loads in samples, which is a highly relevant issue for determining viral persistence in as-yet-unknown tissue reservoirs in individuals suffering from post-COVID conditions or long COVID.

Topics & Concepts

Subgenomic mRNADigital polymerase chain reactionViral loadVirologyBiologyReverse transcription polymerase chain reactionReal-time polymerase chain reactionReverse transcriptaseSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2)CoronavirusRNAViral replicationVirusCoronavirus disease 2019 (COVID-19)Polymerase chain reactionGeneMedicineMessenger RNADiseaseInfectious disease (medical specialty)GeneticsPathologySARS-CoV-2 detection and testingSARS-CoV-2 and COVID-19 ResearchRespiratory viral infections research