Objective To investigate the diagnostic values of ultrasound (US) -guided fine-needle aspiration-thyroglobulin (FNA-Tg), US-guided fine-needle aspiration cytology (FNAC) and their combination in the diagnosis of cervical lymph node metastasis from papillary thyroid carcinoma (PTC). Methods Retrospective collection of 70 patients with suspected lymph node metastasis of PTC, the US-guided FNA was performed on the suspicious lymph node, followed by FNA-Tg and FNAC testing. The receiver operating characteristic curves were plotted taking postoperative pathological results as the gold standard, and the optimal cut-off value of FNA-Tg was calculated. Meanwhile, the Kappa tests were applied to compare the consistency of various methods with the gold standard, respectively. Furthermore, the diagnostic efficacy of each method for PTC lymph node metastasis was calculated separately. Results A total of 71 lymph nodes from 70 patients were analyzed, postoperative pathological examination confirmed 40 lymph nodes as metastatic and 31 as non-metastatic from PTC. The areas under the curve of FNA-Tg, FNAC and their combination in diagnosing lymph nodes metastasis from PTC were 0.993, 0.834 and 0.971, with an optimal cutoff value of 115 ng/mL for FNA-Tg. The sensitivity, specificity, accuracy, PPV, NPV and kappa value for FNA-Tg and FNAC in diagnosing PTC metastatic lymph node were 95%, 100%, 97%, 100%, 94%, 0.943, and 70%, 97%, 83%, 97%, 71%, 0.642, respectively. When used in combination, the corresponding values were 97.5%, 97%, 97%, 97.5%, 97% and 0.943. Conclusions The diagnostic efficacy of FNA-Tg and the combination of FNA-Tg and FNAC for lymph node metastasis from PTC, as well as their consistency with postoperative pathology, were superior to that of FNAC alone. The combined application of FNA-Tg and FNAC improved both the sensitivity and the NPV compared to FNA-Tg alone.