Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR
Top Cited Papers
Open Access
- 23 September 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 11 (1), 1-12
- https://doi.org/10.1038/s41467-020-18611-5
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.Funding Information
- Vetenskapsrådet (2017-01723)
- Ragnar Söderbergs stiftelse (M16/17)
- Science for Life Laboratory (2020.0182)
- Knut och Alice Wallenbergs Stiftelse (2020.0182)
- SciLifeLab/KAW national COVID-19 research program project grant
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