SARS-CoV-2 Quantitation from Plasma

Total nucleic acids were extracted from 200 µl of plasma using the MagNA Pure 96 system (Roche, Penzberg, Germany) and eluted in 50 µl. SARS-CoV-2 RNA was detected from 5 µL eluate in a 25 µL reaction mix on a QuantStudio™ 7 PCR system (Thermofisher Scientific, Waltham, MA, USA) in duplicate reactions, according to the method published by Corman et al.^{18 (link)}. SARS-CoV-2 RNA quantitation was calculated using a serial dilution of a synthetic Wuhan coronavirus 2019 E gene RNA control, provided by the European Virus Archive Global (EVAg). The limit of detection (LoD) and limit of quantitation (LoQ) of the assay was 2.29 and 2.70 log₁₀ RNA copies/mL plasma, respectively.