We electrophysiologically evaluated the autonomic function (AF) in a consecutive series of patients with beta-thalassemia and in normal individuals. Six quantitative autonomic function tests (AFTs) were used: tilt test, hand grip test and sympathetic skin response for sympathetic function; R-R interval, inspiration-expiration difference and 30/15 ratio for parasympathetic function. The prevalence of impaired AF was higher in beta-thalassemia patients (13%, n = 5) than in control subjects (0%, n = 0; p = 0.026). Subclinical autonomic dysfunction appeared to be more prevalent in beta-thalassemia patients compared to controls in our series. Further independent validation of this finding is required in larger cohorts of beta-thalassemia patients.