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....

Full description

Saved in:
Bibliographic Details
Published in:Nature communications Vol. 11; no. 1; pp. 4812 - 12
Main Authors: 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
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 23.09.2020
Nature Portfolio
Subjects:
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
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
Bibliography: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