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The expanded quantitative urine culture protocol was used to compare the microbial abundance and diversity of voided urines obtained using a standard urine collection or using the Peezy midstream device versus paired periurethral specimens.Sixty-two female participants were assigned to 1 of 3 cohorts. One cohort used a standard clean catch midstream urine protocol that included a castile soap wipe, the second cohort used a Peezy midstream collection device with castile soap wipe and the third used the Peezy device without a castile soap wipe. Each participant watched a video that detailed the collection method. Before using the castile soap wipe, a periurethral swab was obtained to measure periurethral microbial abundance. Demographics and pelvic floor symptoms were assessed by validated questionnaires. Microbes were detected using expanded quantitative urine culture. Diversity within each sample was analyzed using alpha diversity measures.Paired periurethral and urine samples for each woman were analyzed and compared for species abundance, richness, and diversity. Bacterial profiles of Peezy-collected urines differed significantly by multiple diversity indices and had significantly reduced colony-forming units compared to paired periurethral swabs. In contrast, within the standard clean catch cohort, voided urine had higher abundance and richness than paired periurethral swabs.Compared with standard clean catch method, the Peezy urine collection device with and without the castile soap wipe resulted in urine with lower bacterial abundance that was distinct from the periurethra. Voided urine collected by Peezy may reduce postbladder microbial contribution.