One-step RNA extraction for RT-qPCR detection of 2019-nCoV

Monica Sentmanat; Evguenia Kouranova; Xiaoxia Cui

We presented here an improvement to the standard 2019-nCoV RT-qPCR test. We modified the RNA extraction step by using a one-step lysis buffer, bypassing the use of column purification. Our indicators for adequate assay performance are consistent with CDC assay interpretation guidelines, the detection of a Ct level of <40 for 2019-nCoV positive control counterparts and for RNaseP reference signal for positive controls and experimental samples 4 . The direct lysis takes less than 20 min to process samples ready for RT-qPCR and can be easily scaled up to a 96-well format and obtain higher throughput. The cost of the lysis buffer is much lower than that of a column purification kit. The lysis only requires a regular PCR machine and does not need a centrifuge or a manifold, as column purification requires.