Comparison of real‐time and droplet digital PCR to detect and quantify SARS‐CoV‐2 RNA in plasma

Background The presence of SARS‐CoV‐2 RNA in plasma has been linked to disease severity and mortality. We compared RT‐qPCR to droplet digital PCR (ddPCR) to detect SARS‐CoV‐2 RNA in plasma from COVID‐19 patients (mild, moderate, and critical disease). Methods The presence/concentration of SARS‐CoV‐2...

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Published in:European Journal of Clinical Investigation 2021-06, Vol.51 (6), p.e13501-n/a
Main Authors: Tedim, Ana P., Almansa, Raquel, Domínguez‐Gil, Marta, González‐Rivera, Milagros, Micheloud, Dariela, Ryan, Pablo, Méndez, Raúl, Blanca‐López, Natalia, Pérez‐García, Felipe, Bustamante, Elena, Gómez, José Manuel, Doncel, Cristina, Trapiello, Wysali, Kelvin, Alyson A., Booth, Ryan, Ostadgavahi, Ali Toloue, Oneizat, Ruth, Puertas, Carolina, Barbé, Ferrán, Ferrer, Ricard, Menéndez, Rosario, Bermejo‐Martin, Jesús F, Eiros, José María, Kelvin, David J, Torres, Antoni
Format: Article
Language:English
Subjects:
COVID-19
ddPCR
Droplets
Original
Plasma
Ribonucleic acid
RNA
RNAemia
RT‐qPCR
SARS‐CoV‐2
Severe acute respiratory syndrome
Severe acute respiratory syndrome coronavirus 2
Statistical analysis
Statistics
Viral diseases
viral RNA load
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Summary:Background The presence of SARS‐CoV‐2 RNA in plasma has been linked to disease severity and mortality. We compared RT‐qPCR to droplet digital PCR (ddPCR) to detect SARS‐CoV‐2 RNA in plasma from COVID‐19 patients (mild, moderate, and critical disease). Methods The presence/concentration of SARS‐CoV‐2 RNA in plasma was compared in three groups of COVID‐19 patients (30 outpatients, 30 ward patients and 30 ICU patients) using both RT‐qPCR and ddPCR. Plasma was obtained in the first 24h following admission, and RNA was extracted using eMAG. ddPCR was performed using Bio‐Rad SARS‐CoV‐2 detection kit, and RT‐qPCR was performed using GeneFinder™ COVID‐19 Plus RealAmp Kit. Statistical analysis was performed using Statistical Package for the Social Science. Results SARS‐CoV‐2 RNA was detected, using ddPCR and RT‐qPCR, in 91% and 87% of ICU patients, 27% and 23% of ward patients and 3% and 3% of outpatients. The concordance of the results obtained by both methods was excellent (Cohen's kappa index = 0.953). RT‐qPCR was able to detect 34/36 (94.4%) patients positive for viral RNA in plasma by ddPCR. Viral RNA load was higher in ICU patients compared with the other groups (P 85%). RT‐qPCR was as useful as ddPCR to detect and quantify SARS‐CoV‐2 RNAemia in plasma.
ISSN:0014-2972
1365-2362
DOI:10.1111/eci.13501