16s RNA-seq of Homo sapiens: adult female fecal sample
16S rRNA sequencing was performed on fecal samples from 32 Hunner-type IC/BPS patients and 30 healthy controls, following previously established protocols. The V3V4 hypervariable regions of the bacterial 16S rRNA gene were amplified using primers 338F and 806R on a GeneAmp 9700 PCR system. Sequencing was conducted on an Illumina MiSeq PE300 platform (Illumina, San Diego, USA) according to the standard protocols of Majorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China). Alpha diversity indices (Chao, ACE, Shannon, Simpson) were calculated to assess within-sample diversity, and the coverage index was used to evaluate sequencing depth. Beta diversity (inter-individual variation) was analyzed via principal coordinates analysis (PCoA) and the Adonis test. Differentially abundant taxa between groups were identified using Linear Discriminant Analysis Effect Size (LEfSe).
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