16S rRNA sequencing and conventional culture provide complementary information in hospitalized patients with chronic lower-limb wounds.

Chronic lower-limb wounds are polymicrobial and difficult to characterize using conventional culture alone. We compared baseline wound microbiota assessed by 16S rRNA sequencing and conventional culture in 62 hospitalized adults with chronic lower-limb wounds and examined associations with subsequent length of stay. Sequencing detected a mean of 11.62 taxa per sample versus 2.24 by culture, with an overall concordance of 51.35%. In sequencing-based analyses, alpha diversity was not associated with length of stay, whereas beta diversity differed significantly according to hospitalization duration (PERMANOVA, R² = 0.030, p = 0.005), and six genera were associated with length of stay. Conventional culture showed no significant association with length of stay. No significant associations were found between sequencing-derived baseline microbiota and wound thermal parameters or 12-week healing. These findings indicate that 16S rRNA sequencing and conventional culture provide distinct yet complementary views of chronic wound microbiology.