- Wise, Emma;
- Márquez, Sully;
- Mellors, Jack;
- Paz, Verónica;
- Atkinson, Barry;
- Gutierrez, Bernardo;
- Zapata, Sonia;
- Coloma, Josefina;
- Pybus, Oliver;
- Jackson, Simon;
- Trueba, Gabriel;
- Fejer, Gyorgy;
- Logue, Christopher;
- Pullan, Steven
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.