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Analytical sensitivity and efficiency comparisons of SARS-CoV-2 RT-qPCR primer-probe sets.
- Source :
-
Nature microbiology [Nat Microbiol] 2020 Oct; Vol. 5 (10), pp. 1299-1305. Date of Electronic Publication: 2020 Jul 10. - Publication Year :
- 2020
-
Abstract
- The recent spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) exemplifies the critical need for accurate and rapid diagnostic assays to prompt clinical and public health interventions. Currently, several quantitative reverse transcription-PCR (RT-qPCR) assays are being used by clinical, research and public health laboratories. However, it is currently unclear whether results from different tests are comparable. Our goal was to make independent evaluations of primer-probe sets used in four common SARS-CoV-2 diagnostic assays. From our comparisons of RT-qPCR analytical efficiency and sensitivity, we show that all primer-probe sets can be used to detect SARS-CoV-2 at 500 viral RNA copies per reaction. The exception for this is the RdRp-SARSr (Charité) confirmatory primer-probe set which has low sensitivity, probably due to a mismatch to circulating SARS-CoV-2 in the reverse primer. We did not find evidence for background amplification with pre-COVID-19 samples or recent SARS-CoV-2 evolution decreasing sensitivity. Our recommendation for SARS-CoV-2 diagnostic testing is to select an assay with high sensitivity and that is regionally used, to ease comparability between outcomes.
- Subjects :
- Betacoronavirus isolation & purification
COVID-19
COVID-19 Testing
Clinical Laboratory Techniques statistics & numerical data
Coronavirus Infections epidemiology
Genetic Variation
Genome, Viral
Humans
Molecular Probe Techniques statistics & numerical data
Pandemics
Pneumonia, Viral epidemiology
RNA genetics
RNA Probes genetics
Reverse Transcriptase Polymerase Chain Reaction statistics & numerical data
SARS-CoV-2
Sensitivity and Specificity
Betacoronavirus genetics
Clinical Laboratory Techniques methods
Coronavirus Infections diagnosis
Coronavirus Infections virology
Pneumonia, Viral diagnosis
Pneumonia, Viral virology
RNA, Viral analysis
RNA, Viral genetics
Reverse Transcriptase Polymerase Chain Reaction methods
Subjects
Details
- Language :
- English
- ISSN :
- 2058-5276
- Volume :
- 5
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Nature microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 32651556
- Full Text :
- https://doi.org/10.1038/s41564-020-0761-6