Elevated urinary levels of the renal tubular enzyme, N-acetyl-beta-D-glucosaminidase (NAG), have been shown to be associated with reversible tubular damage and, therefore, may serve as an indicator of tubular damage in the setting of presumed obstruction uropathy. This study compares urinary NAG levels in children with apparent upper tract obstruction with normal children to assess the sensitivity of this assay for the detection of possible renal tubular damage. The study included 40 children 3 weeks to 16 years old with unilateral ureteropelvic junction obstruction (30) or primary obstructive megaureter (10). Urine was obtained from the bladder in all children and from the renal pelvis or ureter in 30 patients at surgery. Pelvic and ureteral urinary NAG levels were consistently higher than bladder levels. In patients with ureteropelvic junction obstruction NAG levels were 7 times higher than normal (76 units per mg., p < 0.0001) and 3 times higher than normal in patients with obstructive megaureter (29 units per mg., p < 0.001). The mean bladder urinary NAG levels in patients with ureteropelvic junction obstruction (17.6 units per mg. creatinine, standard error of mean 2.01, p < 0.001) and megaureters (19.2 units per mg. creatinine, standard error of mean 3.6, p < 0.049) were elevated above control patients (10.6 units per mg. creatinine, standard error of mean 1.02). Elevated urinary NAG levels in the renal pelvis, ureter and bladder may be helpful in identifying upper tract obstruction, which if left untreated, might cause progressive renal deterioration.