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Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR
- Source :
- Nature Communications, Nature Communications, Vol 11, Iss 1, Pp 1-12 (2020)
- Publication Year :
- 2020
-
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.<br />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.
- Subjects :
- 0301 basic medicine
Hot Temperature
General Physics and Astronomy
Viral Plaque Assay
0302 clinical medicine
COVID-19 Testing
Medicine
lcsh:Science
Virus quantification
Multidisciplinary
Reverse Transcriptase Polymerase Chain Reaction
Reverse transcription polymerase chain reaction
Benchmarking
Real-time polymerase chain reaction
030220 oncology & carcinogenesis
RNA, Viral
RNA extraction
Coronavirus Infections
Validation study
Coronavirus disease 2019 (COVID-19)
Science
Sample (material)
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Pneumonia, Viral
Computational biology
Sensitivity and Specificity
General Biochemistry, Genetics and Molecular Biology
Article
03 medical and health sciences
Betacoronavirus
Viral genetics
Humans
In patient
Pandemics
DNA Primers
Sweden
business.industry
Clinical Laboratory Techniques
SARS-CoV-2
Extraction (chemistry)
Laboratory techniques and procedures
COVID-19
General Chemistry
Virology
030104 developmental biology
Viral infection
lcsh:Q
business
Subjects
Details
- ISSN :
- 20411723
- Volume :
- 11
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Nature communications
- Accession number :
- edsair.doi.dedup.....d369be28428dc53a027290091c24ceac