Real-time RT-PCR for the detection and quantitation of Oropouche virus

• Published in: Diagnostic microbiology and infectious disease 2020-01, Vol.96 (1), p.114894-114894, Article 114894
• Main Authors: Rojas, Alejandra, Stittleburg, Victoria, Cardozo, Fátima, Bopp, Nathen, Cantero, César, López, Sanny, Bernal, Cynthia, Mendoza, Laura, Aguilar, Patricia, Pinsky, Benjamin A., Guillén, Yvalena, Páez, Malvina, Waggoner, Jesse J.
• Format: Article
• Language: English
• Subjects: Adult; Bunyaviridae Infections - diagnosis; Bunyaviridae Infections - virology; Female; Humans; Limit of Detection; Male; Middle Aged; Oropouche virus; Orthobunyavirus; Orthobunyavirus - isolation & purification; Paraguay; Quantitative real-time PCR; Real-Time Polymerase Chain Reaction; Reverse transcriptase PCR; RNA, Viral - isolation & purification; Sensitivity and Specificity
• ISSN: 0732-8893; 1879-0070
• DOI: 10.1016/j.diagmicrobio.2019.114894

Oropouche virus (OROV) causes an acute, systemic febrile illness, and in certain regions of South America, this represents the second most common human arboviral infection after dengue virus. A new real-time RT-PCR was developed for OROV and reassortant species. The new OROV rRT-PCR proved linear across 6–7 orders of magnitude with a lower limit of 95% detection of 5.6–10.8 copies/μL. Upon testing dilutions of OROV and Iquitos virus reference genomic RNA, all dilutions with >10 copies/μL were detected in both the OROV rRT-PCR and a comparator molecular assay, but the OROV rRT-PCR detected more samples with ≤10 copies/μL (8/14 vs 0/13, respectively, P = 0.002). In a set of 100 acute-phase clinical samples from Paraguay patients with a suspected arboviral illness, no patients tested positive for OROV RNA using either assay. The OROV rRT-PCR provides a sensitive molecular assay for the study of this important yet neglected tropical arboviral infection.