Oropouche virus cases identified in Ecuador using an optimised qRT-PCR informed by metagenomic sequencing.

Oropouche virus (OROV) is responsible for outbreaks of Oropouche fever in parts of South America. We recently identified and isolated OROV from a febrile Ecuadorian patient, however, a previously published qRT-PCR assay did not detect OROV in the patient sample. A primer mismatch to the Ecuadorian O...

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Main Authors: Emma L Wise (Author), Sully Márquez (Author), Jack Mellors (Author), Verónica Paz (Author), Barry Atkinson (Author), Bernardo Gutierrez (Author), Sonia Zapata (Author), Josefina Coloma (Author), Oliver G Pybus (Author), Simon K Jackson (Author), Gabriel Trueba (Author), Gyorgy Fejer (Author), Christopher H Logue (Author), Steven T Pullan (Author)
Format: Book
Published: Public Library of Science (PLoS), 2020-01-01T00:00:00Z.
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Summary:Oropouche virus (OROV) is responsible for outbreaks of Oropouche fever in parts of South America. We recently identified and isolated OROV from a febrile Ecuadorian patient, however, a previously published qRT-PCR assay did not detect OROV in the patient sample. A primer mismatch to the Ecuadorian OROV lineage was identified from metagenomic sequencing data. We report the optimisation of an qRT-PCR assay for the Ecuadorian OROV lineage, which subsequently identified a further five cases in a cohort of 196 febrile patients. We isolated OROV via cell culture and developed an algorithmically-designed primer set for whole-genome amplification of the virus. Metagenomic sequencing of the patient samples provided OROV genome coverage ranging from 68-99%. The additional cases formed a single phylogenetic cluster together with the initial case. OROV should be considered as a differential diagnosis for Ecuadorian patients with febrile illness to avoid mis-diagnosis with other circulating pathogens.
Item Description:1935-2727
1935-2735
10.1371/journal.pntd.0007897