Metagenomic sequencing of human urinary tract metagenome: female UTI urine sample
Total DNA was extracted from urine samples using protocols optimized for low-biomass specimens. Bacterial community profiling was performed via 16S rRNA gene amplicon sequencing targeting the V3V4 hypervariable regions. Sequencing was conducted on an Illumina NextSeq 2000 platform using a 600-cycle kit, generating paired-end reads. Raw reads were quality-filtered and denoised to generate amplicon sequence variants (ASVs), with chimeric sequences removed. Taxonomy was assigned using the SILVA NR99 reference database (v138.1) at phylum-to-genus level, retaining unassigned labels when confidence thresholds were not met. Samples with low sequencing depth (<5,000 non-chimeric reads) were excluded from downstream analyses.