Detection of SARS-CoV-2 RNA by multiplex RT-qPCR.
PLoS Biol
; 18(10): e3000867, 2020 10.
Article
in English
| MEDLINE | ID: covidwho-901993
Preprint
This scientific journal article is probably based on a previously available preprint. It has been identified through a machine matching algorithm, human confirmation is still pending.
See preprint
This scientific journal article is probably based on a previously available preprint. It has been identified through a machine matching algorithm, human confirmation is still pending.
See preprint
ABSTRACT
The current quantitative reverse transcription PCR (RT-qPCR) assay recommended for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing in the United States requires analysis of 3 genomic targets per sample 2 viral and 1 host. To simplify testing and reduce the volume of required reagents, we devised a multiplex RT-qPCR assay to detect SARS-CoV-2 in a single reaction. We used existing N1, N2, and RP primer and probe sets by the Centers for Disease Control and Prevention, but substituted fluorophores to allow multiplexing of the assay. The cycle threshold (Ct) values of our multiplex RT-qPCR were comparable to those obtained by the single assay adapted for research purposes. Low copy numbers (≥500 copies/reaction) of SARS-CoV-2 RNA were consistently detected by the multiplex RT-qPCR. Our novel multiplex RT-qPCR improves upon current single diagnostics by saving reagents, costs, time, and labor.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Pneumonia, Viral
/
Reagent Kits, Diagnostic
/
RNA, Viral
/
Coronavirus Infections
/
Clinical Laboratory Techniques
/
Reverse Transcriptase Polymerase Chain Reaction
/
Multiplex Polymerase Chain Reaction
/
Betacoronavirus
Type of study:
Diagnostic study
/
Observational study
/
Prognostic study
Limits:
Humans
Country/Region as subject:
North America
Language:
English
Journal:
PLoS Biol
Journal subject:
Biology
Year:
2020
Document Type:
Article
Affiliation country:
JOURNAL.PBIO.3000867
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