Detection of SARS-CoV-2 RNA by multiplex RT-qPCR.
PLoS Biol
; 18(10): e3000867, 2020 10.
Article
en En
| MEDLINE
| ID: mdl-33027248
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.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Neumonía Viral
/
Juego de Reactivos para Diagnóstico
/
ARN Viral
/
Infecciones por Coronavirus
/
Técnicas de Laboratorio Clínico
/
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Reacción en Cadena de la Polimerasa Multiplex
/
Betacoronavirus
Tipo de estudio:
Diagnostic_studies
/
Observational_studies
Límite:
Humans
País/Región como asunto:
America do norte
Idioma:
En
Revista:
PLoS Biol
Asunto de la revista:
BIOLOGIA
Año:
2020
Tipo del documento:
Article
País de afiliación:
Estados Unidos