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Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR

Authors :
Antonio Lentini
Björn Reinius
Hamzah Safari
Ioanna Smyrlaki
Antonio Gigliotti Rothfuchs
Björn Högberg
Shaman Muradrasoli
Johan Aarum
Jan Albert
Nuno Rufino de Sousa
Natali Papanicolaou
Martin Ekman
Martin Vondracek
Source :
Nature Communications, Nature Communications, Vol 11, Iss 1, Pp 1-12 (2020)
Publication Year :
2020

Abstract

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in patient samples, but RNA extraction constitutes a major bottleneck in current testing. Methodological simplification could increase diagnostic availability and efficiency, benefitting patient care and infection control. Here, we describe methods circumventing RNA extraction in COVID-19 testing by performing RT-PCR directly on heat-inactivated or lysed samples. Our data, including benchmarking using 597 clinical patient samples and a standardised diagnostic system, demonstrate that direct RT-PCR is viable option to extraction-based tests. Using controlled amounts of active SARS-CoV-2, we confirm effectiveness of heat inactivation by plaque assay and evaluate various generic buffers as transport medium for direct RT-PCR. Significant savings in time and cost are achieved through RNA-extraction-free protocols that are directly compatible with established PCR-based testing pipelines. This could aid expansion of COVID-19 testing.<br />SARS-CoV-2 infection is widely diagnosed by RT-PCR, but RNA extraction is a bottleneck for fast and cheap diagnosis. Here, the authors develop protocols to perform RT-PCR directly on heat-inactivated subject samples or samples lysed with readily available detergents and benchmark performance against 597 clinically diagnosed patient samples.

Details

ISSN :
20411723
Volume :
11
Issue :
1
Database :
OpenAIRE
Journal :
Nature communications
Accession number :
edsair.doi.dedup.....d369be28428dc53a027290091c24ceac