Purpose: The oxygen uptake efficiency slope (OUES) has been proposed as an ‘effort-independent’ measure of cardiopulmonary exercise capacity, which could be used as an alternative measurement for peak oxygen uptake (VO2peak) in populations unable or unwilling to perform maximal exercise. The aim of the current study was to investigate the validity of the OUES in children with cystic fibrosis (CF). Methods: Exercise data of 22 children with CF and mild to moderate airflow obstruction were analyzed and compared with exercise data of 22 healthy children. The OUES was calculated using data up to three different relative exercise intensities, namely 50%, 75%, and 100% of the total exercise duration, and normalized for body surface area (BSA). Results: Only the OUES/BSA using the first 50% of the total exercise duration was significantly different between the groups. OUES/BSA values determined at different exercise intensities differed significantly within patients with CF and correlated only moderately with VO2peak and the ventilatory threshold. Conclusion: The OUES is not a valid submaximal measure of cardiopulmonary exercise capacity in children with mild to moderate CF, due to its limited distinguishing properties, its nonlinearity throughout progressive exercise, and its moderate correlation with VO2peak and the ventilatory threshold.