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

Alejandra Rojas, Victoria Stittleburg, Fátima Cardozo, Nathen Bopp, César Cantero, Sanny López, Cynthia Bernal, Laura Mendoza, Patricia Aguilar, Benjamin A. Pinsky, Yvalena Guillén, Malvina Páez, Jesse J. Waggoner

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Article number114894
JournalDiagnostic Microbiology and Infectious Disease
Volume96
Issue number1
DOIs
StatePublished - Jan 2020

Fingerprint

Real-Time Polymerase Chain Reaction
Viruses
Polymerase Chain Reaction
Reassortant Viruses
Paraguay
Dengue Virus
South America
RNA Viruses
Infection
Limit of Detection
Fever
RNA

Keywords

  • Oropouche virus
  • Orthobunyavirus
  • Quantitative real-time PCR
  • Reverse transcriptase PCR

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

Cite this

Rojas, A., Stittleburg, V., Cardozo, F., Bopp, N., Cantero, C., López, S., ... Waggoner, J. J. (2020). Real-time RT-PCR for the detection and quantitation of Oropouche virus. Diagnostic Microbiology and Infectious Disease, 96(1), [114894]. https://doi.org/10.1016/j.diagmicrobio.2019.114894

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

In: Diagnostic Microbiology and Infectious Disease, Vol. 96, No. 1, 114894, 01.2020.

Research output: Contribution to journalArticle

Rojas, A, Stittleburg, V, Cardozo, F, Bopp, N, Cantero, C, López, S, Bernal, C, Mendoza, L, Aguilar, P, Pinsky, BA, Guillén, Y, Páez, M & Waggoner, JJ 2020, 'Real-time RT-PCR for the detection and quantitation of Oropouche virus', Diagnostic Microbiology and Infectious Disease, vol. 96, no. 1, 114894. https://doi.org/10.1016/j.diagmicrobio.2019.114894
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. / Real-time RT-PCR for the detection and quantitation of Oropouche virus. In: Diagnostic Microbiology and Infectious Disease. 2020 ; Vol. 96, No. 1.
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