BACKGROUND The purpose of this study was to prospectively evaluate parathyroid localization using technetium-99m-sestamibi (MIBI). METHODS Technetium-99m-sestamibi scintigraphy was performed in 124 patients with hyperparathyroidism and the results were correlated with serum calcium and parathyroid hormone (PTH) levels, weight, location, and pathology of parathyroid tissue, and associated thyroid abnormalities. RESULTS Hyperparathyroidism was primary in 118 patients, secondary in four patients, and tertiary in two patients. The parathyroid pathology was a solitary adenoma in 95 patients (77 percent), double adenoma in five (4 percent), hyperplasia in 14 (11 percent), carcinoma in one (1 percent), and unconfirmed in nine (7 percent) who underwent noncurative parathyroidectomy. Associated thyroid disease was present in 29 (23 percent) patients. Fourteen patients (11 percent) had undergone previous parathyroid exploration. The mean calcium level was 11.4 +/- 0.8 mg/dL (range, 8.3 to 13.7 mg/dL) and the mean adjusted PTH level was 395 +/- 702 (range, 70 to 4,331). The sensitivity and positive predictive value of MIBI scintigraphy were 81 and 89 percent, respectively, in patients with a solitary adenoma and 37 and 100 percent, respectively, in patients with multiglandular disease. The mean adjusted PTH level was higher in patients with true-positive scans compared with false-negative scans (440 +/- 628 compared with 243 +/- 499, p > 0.05). The mean adenoma weight was 1,877 +/- 3,212 mg in patients with a true-positive scan compared with 485 +/- 296 mg with a false-negative scan (p > 0.05). CONCLUSIONS The sensitivity and positive predictive value of MIBI scintigraphy is comparable to or better than the results reported for other localization procedures. Its lack of sensitivity for detection of multiglandular disease precludes its use in lieu of routine bilateral neck exploration in the management of patients with hyperparathyroidism.