Objective To investigate the value of ultrasonic shear wave elastography (SWE) combined with serum midkine (MK), thymidine kinase 1 (TK1), and the thyrotropin and thyroglobulin (TSH/Tg) ratio in the differential diagnosis of benign and malignant thyroid nodules. Methods The clinical data of 129 patients with thyroid nodules (129 nodules) who were admitted to our hospital from May 2020 to May 2022 were retrospectively selected. According to postoperative pathological results or puncture results, they were divided into benign group with 98 cases and malignant group with 31 cases, all of which underwent ultrasonic SWE examination and serum MK, TK1, TSH and Tg detection. SWE parameters [minimum modulus of elasticity (Emin), maximum modulus of elasticity (Emax) and average modulus of elasticity (Emean)] in the two groups were compared, and the serum MK, TK1 and TSH/Tg ratio in the two groups were compared. Receiver operating characteristic (ROC) curve was used to analyze the differential diagnosis value of SWE parameters combined with serum MK, TK1 and TSH/Tg ratio for benign and malignant thyroid nodules. Results Compared with the benign group, the Emax, Emin and Emean value and the serum MK, TK1 and TSH/Tg ratio in the malignant group were significantly increased (P＜0.05). The area under curve (AUC), sensitivity and specificity of SWE parameter combined with serum MK, TK1 and TSH/Tg ratio were 0.893, 83.92% and 89.82%, respectively. The value of SWE parameter in the differential diagnosis of benign and malignant thyroid nodules was significantly better than that of each indicator alone. Conclusion The SWE parameters Emax, Emin, Emean values and serum MK, TK1, TSH/Tg ratio are significantly different in thyroid nodules of different properties. Their combined detection is of high value in the differential diagnosis of benign and malignant thyroid nodules, which is more advantageous than the single application of various indicators.