Homo sapiens Metagenome for viral nucleic acid detection in bone marrow specimens

In 2016, after informed consent was gained, as approved by the Medical Ethics Committee of Jiangsu University, 24 bone marrow specimens were collected from 24 hematologic patients. For library preparation, 100 µL of each specimen was pipetted and randomly pooled into 4 pools each including 6 samples. The 4 sample pools were thawed three times on dry-ice, the supernatants were then collected after centrifugation (10 min, 15,000 × g). Library preparation was performed as previously described. Briefly, pooled supernatants were filtered through a 0.45-mm filter (Millipore), and 200 µL of each pool was then subjected to a mixture of nuclease enzymes. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit (Qiagen) and 4 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. Bioinformatics analysis was performed according to a previous study