Comparison of real-time and droplet digital PCR to detect and quantify SARS-CoV-2 RNA in plasma.
COVID-19
RNAemia
RT-qPCR
SARS-CoV-2
ddPCR
viral RNA load
Journal
European journal of clinical investigation
ISSN: 1365-2362
Titre abrégé: Eur J Clin Invest
Pays: England
ID NLM: 0245331
Informations de publication
Date de publication:
Jun 2021
Jun 2021
Historique:
revised:
13
01
2021
received:
22
12
2020
accepted:
25
01
2021
pubmed:
30
1
2021
medline:
16
6
2021
entrez:
29
1
2021
Statut:
ppublish
Résumé
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). 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. 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 < .001), by both ddPCR and RT-qPCR. AUC analysis revealed Ct values (RT-qPCR) and viral RNA load values (ddPCR) can similarly differentiate between patients admitted to wards and to the ICU (AUC of 0.90 and 0.89, respectively). Both methods yielded similar prevalence of RNAemia between groups, with ICU patients showing the highest (>85%). RT-qPCR was as useful as ddPCR to detect and quantify SARS-CoV-2 RNAemia in plasma.
Sections du résumé
BACKGROUND
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
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
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 < .001), by both ddPCR and RT-qPCR. AUC analysis revealed Ct values (RT-qPCR) and viral RNA load values (ddPCR) can similarly differentiate between patients admitted to wards and to the ICU (AUC of 0.90 and 0.89, respectively).
CONCLUSION
CONCLUSIONS
Both methods yielded similar prevalence of RNAemia between groups, with ICU patients showing the highest (>85%). RT-qPCR was as useful as ddPCR to detect and quantify SARS-CoV-2 RNAemia in plasma.
Identifiants
pubmed: 33512013
doi: 10.1111/eci.13501
pmc: PMC7995030
doi:
Substances chimiques
RNA, Viral
0
Types de publication
Comparative Study
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e13501Subventions
Organisme : European Society of Clinical Microbiology and Infectious Diseases
ID : Research Grant 2020 APT
Organisme : CIHR
ID : OV2 - 170357
Pays : Canada
Organisme : Instituto de Salud Carlos III
ID : CD018/0123
Organisme : Instituto de Salud Carlos III
ID : CIBERES 06/06/0028
Organisme : Instituto de Salud Carlos III
ID : COV20/00110
Organisme : CIHR
ID : OV2 - 170357
Pays : Canada
Informations de copyright
© 2021 Stichting European Society for Clinical Investigation Journal Foundation. Published by John Wiley & Sons Ltd.
Références
Crit Care. 2020 Dec 14;24(1):691
pubmed: 33317616
Emerg Microbes Infect. 2020 Dec;9(1):469-473
pubmed: 32102625
J Infect. 2021 Mar;82(3):414-451
pubmed: 33131699
J Hosp Infect. 2020 May;105(1):1-2
pubmed: 32173458
Int J Antimicrob Agents. 2021 Feb;57(2):106249
pubmed: 33259918
Clin Infect Dis. 2020 Nov 5;71(8):1937-1942
pubmed: 32301997
Nat Commun. 2021 Jun 7;12(1):3406
pubmed: 34099652
Eur J Clin Invest. 2021 Jun;51(6):e13501
pubmed: 33512013
J Clin Microbiol. 1990 Mar;28(3):495-503
pubmed: 1691208
Anaesthesia. 2020 Oct;75(10):1340-1349
pubmed: 32602561
Nat Methods. 2013 Oct;10(10):1003-5
pubmed: 23995387
Clin Infect Dis. 2021 Nov 2;73(9):e2890-e2897
pubmed: 32803231
J Clin Microbiol. 2020 Oct 21;58(11):
pubmed: 32817231