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

Refinement of the genome assembly. Orthobunyavirus members are known to possess characteristic short consensus sequences at their termini (5′ and 3′) 24 . In-depth analysis of the NZV sequence revealed that some of the segment termini lacked this consensus sequence. We therefore applied an additional algorithm for de novo assembly using the SPAdes tool. This analysis resulted in contigs that were similar to but longer than those obtained from the Velvet algorithm, and these contigs included the conserved sequences at all termini, which were flanked by short sequences that were apparently host-derived [25] [26] [27] . To further refine and verify the sequences of the three segments, we realigned the original sequencing reads to the longer sequences obtained from SPAdes and removed low-quality alignments. This procedure resulted in high-quality, manually curated, www.nature.com/scientificreports www.nature.com/scientificreports/ complete genomic sequences of the three segments of NZV. The complete genomic sequences were annotated and deposited in the NCBI GenBank (accession numbers MH018032, MH018033, and MH018034 for the L, M, and S segments, respectively). Comparison of the 5′ and 3′ ends of each of these NZV segments revealed that the novel virus contains a similar consensus sequence as other Orthobunyavirus species (Fig. 3) . These findings support the classification of NZV as an Orthobunyavirus.