Ninety-four children with febrile urinary tract infection were studied prospectively to determine the relationship between vesicoureteral reflux, P-fimbriated Escherichia coli, and acute pyelonephritis, and to evaluate the diagnostic reliability of commonly used clinical and laboratory observations. By using renal scan with dimercaptosuccinic acid labeled with technetium 99m as the standard of reference, we documented acute pyelonephritis in 62 (66%) of 94 patients. Vesicoureteral reflux was demonstrated in 29 (31%) of the total group and in only 23 (37%) of 62 patients with pyelonephritis. Of the 70 E. coli urinary isolates, 48 (69%) were P-fimbriated, including 30 (64%) of 47 isolates from patients with pyelonephritis and 18 (78%) of 23 isolates from patients with normal renal scans. The prevalence of P-fimbriated E. coli in patients with pyelonephritis and vesicoureteral reflux was 46%, compared with 71% in those with pyelonephritis who had no concurrent vesicoureteral reflux (p = 0.222). Multiple clinical and laboratory variables commonly used in the diagnosis of acute pyelonephritis did not adequately predict the presence or absence of parenchymal involvement. These data show the following: (1) Acute pyelonephritis in the absence of demonstrable vesicoureteral reflux is common. (2) Febrile urinary tract infections in children are commonly associated with P-fimbriated E. coli, both in the presence and absence of vesicoureteral reflux. (3) The presence of P fimbriae alone does not fully explain the pathophysiology of renal parenchymal invasion by bacteria in the absence of vesicoureteral reflux. (4) The diagnosis of acute pyelonephritis in children with febrile urinary tract infections on the basis of clinical and laboratory observations is unreliable.