Recent reports have shown that radio-guided surgery with an intraoperative gamma probe (IGP) is useful for identifying parathyroid adenomas. The aim of this study was to evaluate the usefulness of IGP mapping in patients with secondary hyperparathyroidism. Seven patients with secondary hyperparathyroidism underwent technetium-99m methoxyisobutylisonitrile (Tc-99m MIBI) scintigraphy immediately before undergoing surgical resection of all parathyroid tissues. We compared the sensitivity of Tc-99m MIBI scintigraphy alone with that of the combination of Tc-99m MIBI scintigraphy and IGP mapping. The sensitivity of the combination of Tc-99m MIBI scintigraphy and IGP mapping was 100% (28 of 28 lesions) and was significantly higher than that of Tc-99m MIBI scintigraphy alone (71.4%, 20 of 28 lesions). The combination of the Tc-99m MIBI scintigraphy and IGP mapping has increased sensitivity for identifying parathyroid lesions during parathyroidectomy in patients with secondary hyperparathyroidism.