ddPCR for the Detection and Absolute Quantification of Oropouche Virus.
Oropouche virus
RNA
ddPCR
quantification
Journal
Viruses
ISSN: 1999-4915
Titre abrégé: Viruses
Pays: Switzerland
ID NLM: 101509722
Informations de publication
Date de publication:
07 Sep 2024
07 Sep 2024
Historique:
received:
12
08
2024
revised:
03
09
2024
accepted:
06
09
2024
medline:
29
9
2024
pubmed:
28
9
2024
entrez:
28
9
2024
Statut:
epublish
Résumé
Oropouche virus (OROV) is a segmented RNA virus belonging to the genus The ddPCR reaction was assessed as duplex assay using the human housekeeping gene The LoD of the ddPCR performed using 10-fold serial dilution of OROV detected up to 1 cp/µL in all the biological matrices. Compared to the routine molecular diagnostics, the ddPCR assay showed 100% sensitivity for whole blood and serum and 75% for urine, highlighting higher positive rate of ddPCR. We have established a quantitative RNA detection method of OROV with high sensitivity and specificity based on ddPCR. This test is capable of quantitatively monitoring the viral load of OROV and can contribute, in addition to laboratory diagnosis, to shed light on the pathogenesis, filling in the knowledge gaps of this neglected disease and to the vector control programs.
Sections du résumé
BACKGROUND
BACKGROUND
Oropouche virus (OROV) is a segmented RNA virus belonging to the genus
METHODS
METHODS
The ddPCR reaction was assessed as duplex assay using the human housekeeping gene
RESULTS
RESULTS
The LoD of the ddPCR performed using 10-fold serial dilution of OROV detected up to 1 cp/µL in all the biological matrices. Compared to the routine molecular diagnostics, the ddPCR assay showed 100% sensitivity for whole blood and serum and 75% for urine, highlighting higher positive rate of ddPCR.
CONCLUSION
CONCLUSIONS
We have established a quantitative RNA detection method of OROV with high sensitivity and specificity based on ddPCR. This test is capable of quantitatively monitoring the viral load of OROV and can contribute, in addition to laboratory diagnosis, to shed light on the pathogenesis, filling in the knowledge gaps of this neglected disease and to the vector control programs.
Identifiants
pubmed: 39339902
pii: v16091426
doi: 10.3390/v16091426
pii:
doi:
Substances chimiques
RNA, Viral
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Italian Ministry of Health
ID : Fondi Ricerca Corrente L1P3 and L2P14
Organisme : Italian Ministry of Health
ID : Fondi 5 per mille 2021 5M-2021-23684029
Organisme : EU funding within the MUR PNRR
ID : Extended Partnership initiative on Emerging Infectious Diseases (Project no.PE00000007, INF-ACT)