Nodaviridae sp. Genome sequencing
For viral metagenomic analysis, cerebrospinal fluid specimens were collected from patients with encephalitis of unknown etiology in China. The CSF samples were pooled into one specimen pool and thoroughly mixed, resuspended in 1 mL of DPBS, and subjected to three rapid freeze-thaw cycles on dry ice. The supernatants were collected after centrifugation (10 minutes, 15,000 g) and filtered through a 0.45 um filter (Millipore) to remove eukaryotic and bacterial cell-sized particles. A 200 uL aliquot of the supernatant from the pool was treated with a nuclease enzyme mixture to reduce the concentration of free (non-viral encapsidated) nucleic acids. The remaining total nucleic acids were isolated using the QIAamp MinElute Virus Spin Kit according to the manufacturer's protocol. One library was constructed using the Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced on the Illumina NovaSeq platform with a 150 bp paired-end sequencing strategy.