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

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....

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Vydáno v:	Nature communications Ročník 11; číslo 1; s. 4812 - 12
Hlavní autoři:	Smyrlaki, Ioanna, Ekman, Martin, Lentini, Antonio, Rufino de Sousa, Nuno, Papanicolaou, Natali, Vondracek, Martin, Aarum, Johan, Safari, Hamzah, Muradrasoli, Shaman, Rothfuchs, Antonio Gigliotti, Albert, Jan, Högberg, Björn, Reinius, Björn
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	London Nature Publishing Group UK 23.09.2020 Nature Portfolio
Témata:	38/77 38/88 631/326/596/4130 692/699/255/2514 692/700/139/1420 Benchmarking Betacoronavirus - genetics Betacoronavirus - isolation & purification Clinical Laboratory Techniques - methods Clinical Laboratory Techniques - standards Clinical Laboratory Techniques - statistics & numerical data Coronavirus Infections - diagnosis Coronavirus Infections - epidemiology Coronavirus Infections - virology COVID-19 COVID-19 Testing DNA Primers - genetics Hot Temperature Humanities and Social Sciences Humans multidisciplinary Pandemics Pneumonia, Viral - diagnosis Pneumonia, Viral - epidemiology Pneumonia, Viral - virology Reverse Transcriptase Polymerase Chain Reaction - methods Reverse Transcriptase Polymerase Chain Reaction - standards Reverse Transcriptase Polymerase Chain Reaction - statistics & numerical data RNA, Viral - genetics RNA, Viral - isolation & purification SARS-CoV-2 Science Science (multidisciplinary) Sensitivity and Specificity Sweden - epidemiology Viral Plaque Assay - methods Sweden
ISSN:	2041-1723, 2041-1723
On-line přístup:	Získat plný text
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Shrnutí:	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. 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.
Bibliografie:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Undefined-3
ISSN:	2041-1723 2041-1723
DOI:	10.1038/s41467-020-18611-5