In evaluating cervical lymph node (LN) metastasis from papillary thyroid cancer (PTC), ultrasonography (US)-guided fine-needle aspiration biopsy (FNAB) is very important tool. There were limited number of studies about the diagnostic value of thyroglubion measurement in FNAB (FNAB-Tg) in non-thyroidectomized patients. Therefore, in this study, the authors evaluated the role of FNAB-Tg in diagnosing cervical LN metastases in patients with PTC before thyroidectomy. A total 91 suspicious LNs of 68 patients were undergone US-guided FNAB-Tg and cytology. Any FNAB-Tg concentration above 50 ng/ml considered as positive, irrespective of thyroid gland presence. Based on the final pathology, 49 LNs were positive, and the remaining 42 LNs were negative for metastasis. The sensitivity, specificity, and accuracy of FNAB-Tg in thyroidectomized patients were 80.0, 100.0, and 88.9%, respectively. The diagnostic performance of FNAB-Tg was not compromised by the presence of thyroid gland (sensitivity, specificity and accuracy = 95.0, 90.9 and 93.2%, respectively). FNAB-Tg is useful and simple method for the diagnosis of metastatic cervical LNs from PTC. The diagnostic performance of FNAB-Tg was not compromised by the presence of thyroid gland. Therefore, FNAB-Tg could be performed actively for the LN staging of PTC.