tetano
Editor, Senior Moderator
Nat Commun
. 2020 Sep 23;11(1):4812.
doi: 10.1038/s41467-020-18611-5.
Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR
Ioanna Smyrlaki[SUP] 1 [/SUP], Martin Ekman[SUP] 2 [/SUP], Antonio Lentini[SUP] 1 [/SUP], Nuno Rufino de Sousa[SUP] 3 [/SUP], Natali Papanicolaou[SUP] 1 [/SUP], Martin Vondracek[SUP] 2 [/SUP], Johan Aarum[SUP] 2 [/SUP], Hamzah Safari[SUP] 2 [/SUP], Shaman Muradrasoli[SUP] 4 [/SUP], Antonio Gigliotti Rothfuchs[SUP] 3 [/SUP], Jan Albert[SUP] 2 3 [/SUP], Bj?rn H?gberg[SUP] 1 [/SUP], Bj?rn Reinius[SUP] 5 [/SUP]
Affiliations
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.
. 2020 Sep 23;11(1):4812.
doi: 10.1038/s41467-020-18611-5.
Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR
Ioanna Smyrlaki[SUP] 1 [/SUP], Martin Ekman[SUP] 2 [/SUP], Antonio Lentini[SUP] 1 [/SUP], Nuno Rufino de Sousa[SUP] 3 [/SUP], Natali Papanicolaou[SUP] 1 [/SUP], Martin Vondracek[SUP] 2 [/SUP], Johan Aarum[SUP] 2 [/SUP], Hamzah Safari[SUP] 2 [/SUP], Shaman Muradrasoli[SUP] 4 [/SUP], Antonio Gigliotti Rothfuchs[SUP] 3 [/SUP], Jan Albert[SUP] 2 3 [/SUP], Bj?rn H?gberg[SUP] 1 [/SUP], Bj?rn Reinius[SUP] 5 [/SUP]
Affiliations
- PMID: 32968075
- DOI: 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.