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To test whether urethral catheterization (UC) is better than suprapubic bladder aspiration (SPA) as a method for collection of sterile urine in neonates.Thirty-three babies, requiring sterile collection of urine, were randomly assigned to either urethral catheterization (n = 16), median gestation 28 ± 3.9 weeks, median birth weight 968 g (range 650-4100) or SPA (n = 17), median gestation 26 ± 5.6 weeks, median birth weight 926 g (range 771-4070). The primary outcome was success in obtaining urine. Secondary outcomes were complications and urine culture results.Some urine was obtained in 11 (64.7%) babies in the SPA group and in 13 (81.2%) babies in the catheter group. Sufficient urine for analysis (>0.5 ml) was obtained in 10 (58.8%) in the SPA group versus 5 (31.2%) babies in the catheter group. There were more contaminated specimens in the catheter group but this was not significant in this small study.In this small randomized controlled trial urethral catheterization offered no significant advantage over SPA.