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

Ioanna Smyrlaki, Martin Ekman, Antonio Lentini, Nuno Rufino de Sousa, Natali Papanicolaou, Martin Vondracek, Johan Aarum, Hamzah Safari, Shaman Muradrasoli, Antonio Gigliotti Rothfuchs, Jan Albert, Björn Högberg and Björn Reinius ()
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Ioanna Smyrlaki: Karolinska Institutet
Martin Ekman: Karolinska University Hospital
Antonio Lentini: Karolinska Institutet
Nuno Rufino de Sousa: Tumor and Cell Biology, Karolinska Institutet
Natali Papanicolaou: Karolinska Institutet
Martin Vondracek: Karolinska University Hospital
Johan Aarum: Karolinska University Hospital
Hamzah Safari: Karolinska University Hospital
Shaman Muradrasoli: Public Health Agency of Sweden
Antonio Gigliotti Rothfuchs: Tumor and Cell Biology, Karolinska Institutet
Jan Albert: Karolinska University Hospital
Björn Högberg: Karolinska Institutet
Björn Reinius: Karolinska Institutet

Nature Communications, 2020, vol. 11, issue 1, 1-12

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-18611-5

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