Individual variation in sensitivity to glucocorticoids (GCs) poses a dilemma to the clinician. Currently available assays to determine individual sensitivity to GCs either seem imprecise, or they are based on mitogen-activated lymphocytes, although mitogens themselves may affect cellular GC sensitivity. To avoid these disadvantages, we developed an assay based on the GC-induced accumulation of the 51kDa FK506 binding protein (FKBP51) mRNA in unstimulated peripheral blood mononuclear cells (PBMC), measured using real time PCR. Of several family members tested, only FKBP51 transcript levels showed to be GC-inducible. Furthermore, our bioassay was not affected by progesterone, estradiol, and testosterone. Immunological stimulation of PBMC using tetanus toxoid did not affect bioassay results, and isolated T- and B-lymphocytes showed a similar response to GC stimulation. The intra- and inter-assay variations were 10.6 and 15.9%, respectively. Our bioassay confirms previous reports that a wide variation in GC sensitivity exists in the normal population, yet is able to clearly discriminate a patient with familial GC hyposensitivity from controls. Our bioassay may be suitable to assess altered individual GC sensitivity, and the small amount of PBMC needed for a determination makes this assay easily applicable in a pediatric setting.