DIRECT RT-qPCR DETECTION OF SARS-CoV-2 RNA FROM PATIENT NASOPHARYNGEAL SWABS WITHOUT AN RNA EXTRACTION STEP

BioRxiv : the Preprint Server for Biology
Emily A BruceJason W Botten

Abstract

The ongoing COVID-19 pandemic has caused an unprecedented need for rapid diagnostic testing. The Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO) recommend a standard assay that includes an RNA extraction step from a nasopharyngeal (NP) swab followed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to detect the purified SARS-CoV-2 RNA. The current global shortage of RNA extraction kits has caused a severe bottleneck to COVID-19 testing. We hypothesized that SARS-CoV-2 RNA could be detected from NP samples via a direct RT-qPCR assay that omits the RNA extraction step altogether, and tested this hypothesis on a series of blinded clinical samples. The direct RT-qPCR approach correctly identified 92% of NP samples (n = 155) demonstrated to be positive for SARS-CoV-2 RNA by traditional clinical diagnostic RT-qPCR that included an RNA extraction. Thus, direct RT-qPCR could be a front-line approach to identify the substantial majority of COVID-19 patients, reserving a repeat test with RNA extraction for those individuals with high suspicion of infection but an initial negative result. This strategy would drastically ease supply chokepoints of COVID-19 testing and should...Continue Reading

Citations

Sep 30, 2020·Diagnostics·Arunkumar ArumugamSeason Wong
Jul 29, 2020·Science Translational Medicine·Viet Loan Dao ThiSimon Anders
Oct 8, 2020·MSystems·Ceth W ParkerKasthuri Venkateswaran
Oct 20, 2020·Biology Methods and Protocols·Uday KiranKrishnan Harinivas Harshan
Nov 1, 2020·Scientific Reports·Michela DeianaElena Pomari
Dec 1, 2020·Frontiers in Bioengineering and Biotechnology·Dmitriy A VarlamovKonstantin B Ignatov
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BETA
Assay
PCR

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