Homo sapiens Metagenome investigating virome of anogenital warts. Homo sapiens

For viral metagenomic analysis, 110 wart specimens were collected from 110 patients (including 76 male and 34 female) with AGWs who were admitted to the Department of Dermatology and Venerology of the Affiliated Hospital of Jiangsu University from Sep 2016 to Aug 2017. The mean age of study participants was 42.5±15.56 (16-84) years. The average courses were 3.26±3.14 month (1 week to 15 months). The 110 warts were randomly pooled into 11 specimen pools, each including 10 warts. The wart pools were individually homogenized with a mortar and pestle, resuspended in 1 mL DPBS, and then frozen and thawed rapidly three times on dry-ice. The supernatants were then collected after centrifugation (10 min, 15,000 × g). Supernatants were filtered through a 0.45-mm filter (Millipore) to remove eukaryotic- and bacterial cell-sized particles, and 200 uL of supernatant from each pool was then subjected to a mixture of nuclease enzymes to reduce the concentration of free (non-viral encapsidated) nucleic acids[13,14]. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit according to manufacturer's protocol. Eleven libraries were then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the MiSeq Illumina platform with 250 bases paired ends with dual barcoding for each pool.