Identification and genetic characterization of a novel Orthobunyavirus species by a straightforward high-throughput sequencing-based approach

Shifman, Ohad; Cohen-Gihon, Inbar; Beth-Din, Adi; Zvi, Anat; Laskar, Orly; Paran, Nir; Epstein, Eyal; Stein, Dana; Dorozko, Marina; Wolf, Dana; Yitzhaki, Shmuel; Shapira, Shmuel C.; Melamed, Sharon; Israeli, Ofir

The identification and characterization of the novel NZV Orthobunyavirus described in this study demonstrate the power of HTS, combined with bioinformatic tools, to rapidly identify and gain in-depth understanding of unknown and novel viruses in clinical samples. In a scenario of novel viral emergence, straightforward diagnosis, such as that described in this study, could be applied and be of utmost importance and value. www.nature.com/scientificreports www.nature.com/scientificreports/ FCS, MEM-Eagle nonessential amino acids, 2 mM L-glutamine, 0.1 mg/ml streptomycin, 100 units/ml penicillin, 1.25 units/ml nystatin and 10 mM HEPES buffer was added. Six days post infection, when a massive CPE was evident, the culture medium was collected, and cell debris was removed by centrifugation (1700 RCF, 5 min). The collected medium was used to infect fresh Vero monolayers as described above. Five days post infection, when a CPE was observed, the culture medium was collected and used for analyses. Viral titers were determined as pfu/ml using a plaque assay on Vero monolayers as previously described 38 .