Serum levels of the important hormone 1,25-dihydroxyvitamin D (1,25-diOHD, calcitriol) have not been extensively evaluated in patients with cystic fibrosis (CF) during the critical period of skeletal growth and development. This study was a cross-sectional, observational assessment of 25-hydroxyvitamin D (25-OHD, calcidiol) and 1,25-diOHD levels in 54 patients with CF. The patients' ages ranged from 4.9 years to 19.5 years (mean, 11.0 years). Levels were correlated with pulmonary function tests, chest x-ray scores, height and weight Z scores, skinfold percentiles, CF genotype, serum chemistries, and use of a vitamin supplement. Levels were compared with those in more than 160 other pediatric patients living in the same region, and all assays were done in the same laboratory. Despite low-normal levels of the 25-OHD precursor, there was a high prevalence of low (18%) and marginal (18%) levels of 1,25-diOHD. None of the various parameters examined correlated with either 25-OHD or 1,25-diOHD levels. The cause, clinical significance, and treatment of low levels of this important hormone in children with CF warrant further study.