The widespread existence of extended-spectrum β-lactamase (ESBL) producing Escherichia coli (E. coli) has become a critical threat in developed countries. Prediction rule for ESBL producing E. coli is relevant to see patients with suspected urinary tract infection. We collected clinical and laboratory data and constructed multivariate logistic regression models to develop a clinical prediction rule in the derivation cohort with 1185 patients with urine cultures and validated the rule in the validation cohort with 516 patients. ESBL-producing E. coli was found in 185 patients (16%) in the derivation cohort. When assigning 14 points for being female (odds ratio (OR): 4.2), six points for CRP >5 mg/dl (OR: 1.87), and four points for a history of urinary tract infection (OR: 1.52), the area under the curve (AUC) had 0.67 (95% confidence interval (CI): 0.63-0.70) in the derivation cohort and 0.64 (95% CI: 0.59-0.69] in the validation cohort. The developed prediction rule had moderate accuracy to predict ESBL-producing E. coli in patients with suspected urinary tract infection.