A one-enzyme RT-qPCR Assay for SARS-CoV-2, and Procedures for Reagent Production

Given the scale of ongoing COVID-19 pandemic, need for reliable, scalable testing, and likelihood reagent shortages, especially in resource-poor settings, we have developed an RT-qPCR assay that relies on alternative to conventional viral reverse transcriptases, a thermostable transcriptase/DNA polymerase (RTX) (Ellefson et al., 2016). Here show RTX performs comparably other assays sanctioned by CDC validated kit format. We demonstrate two modes use – (i) dye-based require only polymerase, (ii) TaqMan combination Taq DNA polymerases (as exonuclease does not degrade probe). also provide straightforward recipes purification this RTX. anticipate low resource or point-of-need settings researchers could obtain available constructs begin develop their own assays, within whatever regulatory framework exists them.