RT Journal Article
SR Electronic
T1 One-step RNA extraction for RT-qPCR detection of 2019-nCoV
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.02.022384
DO 10.1101/2020.04.02.022384
A1 Sentmanat, Monica
A1 Kouranova, Evguenia
A1 Cui, Xiaoxia
YR 2020
UL http://biorxiv.org/content/early/2020/04/08/2020.04.02.022384.abstract
AB The global outbreak of coronavirus disease 2019 (COVID-19) has placed an unprecedented burden on healthcare systems as the virus spread from the initial 27 reported cases in the city of Wuhan, China to a global pandemic in under three months1. Resources essential to monitoring virus transmission have been challenged with a demand for expanded surveillance. The CDC 2019-nCoV Real-Time Diagnostic Panel uses a real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) consisting of two TaqMan probe and primer sets specific for the 2019-nCoV N gene, which codes for the nucleocapsid structural protein that encapsulates viral RNA, for the qualitative detection of 2019-nCoV viral RNA in respiratory samples. To isolate RNA from respiratory samples, the CDC lists RNA extraction kits from three manufacturers. In anticipation of a limited supply chain of RNA extraction kits and the need for test scalability, we sought to identify alternative RNA extraction methods. Here we show that direct lysis of respiratory samples can be used in place of RNA extraction kits to run the CDC 2019-nCoV Real-Time Diagnostic assay with the additional benefits of higher throughput, lower cost, faster turnaround and possibly higher senitivity and improved saftey.Competing Interest StatementThe authors have declared no competing interest.