Measurement of serum Tg using ultrasensitive assays is proposed to replace TSH-stimulated Tg measurement in the follow-up of differentiated thyroid cancer (DTC). Aim of our study was to verify this possibility using two ultrasensitive Tg assays. We selected 215 DTC patients with undetectable (<1 ng/ml) basal serum Tg at the time of a recombinant human TSH (rhTSH) stimulation. According to standard criteria, 173 (80.4%) patients were considered free of disease, 17 (7.9%) had documented disease and 25 (11.7%) had no evidence of disease but detectable serum rhTSH-stimulated Tg (biochemical disease). The sera of these patients were re-assayed with two commercial ultrasensitive assays and the results were compared with the clinical data. Basal Access and E-Iason Tg assays were able to distinguish patients with persistent disease or free of disease with a sensitivity of 82.3 and 82.3%, specificity of 85.5 and 86.1%, positive predictive value (PPV) of 35.8 and 36.8%, negative predictive value (NPV) of 98 and 98.6%, respectively. With both assays the addition of neck ultrasound to basal Tg increased the sensitivity and the NPV to 100% and decreased the false negative rate to 0%. In patients with detectable basal Tg without evidence of disease, serum Tg converted from detectable to undetectable in about 80% of the cases during 2-yr follow-up. Our study indicates that the combination of neck ultrasound and basal ultrasensitive Tg allows to identify all patients free of disease and can decrease the need for rhTSH stimulation in nearly 80% of the patients.