Background: Controversy surrounds the routine use of urine culture (UC) in febrile neutropenic children. We aimed to evaluate the need for routine urine studies in febrile neutropenic children with cancer. Procedure: We conducted a prospective study of pediatric cancer patients with urinary continence who presented to the emergency department with febrile neutropenia between 2019 and 2021. Epidemiologic data and clinical and laboratory findings were collected. Urinary tract infection (UTI) was defined as urinary signs/symptoms and positive UC with or without pyuria. Study data were analyzed using descriptive statistics. The diagnostic performance of urinalysis was calculated using positive UC as the gold standard. Results: Positive UC was found in seven of the 205 children included (3.4%; 95% CI 1.4-6-9%), 2 presenting urinary symptoms. UTI prevalence was 1.0% (95% CI 0.1-3.5). A 23.8% prevalence of positive UC was found in patients with urinary symptoms and/or history of urinary tract disease (95% CI 8.2-47.2%) as compared to 1.1% of those without symptoms or history (95% CI 0.1-3.9%) (p<0.001). The sensitivity, specificity, negative predictive value, and area under the curve for urinalysis was 16.7% (95% CI 3.0-56.4), 98.4% (95% CI 95.3-99.4), 97.3% (95% CI 93.9-98.9), and 0.65 (95% CI 0.51-0.79), respectively. Conclusions: UTI is an infrequent cause of infection in these patients. Urinalysis is indicated only in febrile neutropenic children with urinary signs/symptoms and in asymptomatic patients with a history of urinary tract disease. When urine is collected, UC should be requested regardless of the result of the urinalysis.