〔Abstract〕 Objective To study the application value of ultrasound combined with serum thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (Tg-Ab) in evaluating the clinical course of Hashimoto thyroiditis (HT). Methods A total of 100 HT patients treated in Guizhou Aerospace Hospital from September 2021 to August 2022 were selected to be included in the observation group, and 100 healthy subjects who underwent physical examination in Guizhou Aerospace Hospital during the same period were selected to be included in the control group. Color Doppler ultrasound and serum TPO-Ab, Tg-Ab examinations were performed in both groups. The serum TPO-Ab, Tg-Ab levels of the two groups were observed. The histopathological grade of postoperative pathology or hollow needle biopsy in the observation group was analyzed, as well as the serum TPO-Ab and Tg-Ab levels of patients with different postoperative histopathological grades in the observation group. To evaluate the accuracy of ultrasonography, serum TPO-Ab, Tg-Ab alone or in combination, the cytopathological grade of HT was evaluated by using postoperative pathology or hollow needle biopsy as the gold standard. Results The levels of serum TPO-Ab and Tg-Ab in the observation group were higher than those in the control group, and the differences were statistically significant (P < 0.05). In the observation group, 58 patients with histopathological grade Ⅰ, 30 patients with histopathological grade Ⅱ and 12 patients with histopathological grade Ⅲ were observed. The levels of serum TPO-Ab and Tg-Ab in grade Ⅰ patients were lower than those in grade Ⅱ and Ⅲ patients, and the levels of serum TPO-Ab and Tg-Ab in grade Ⅱ patients were lower than those in grade Ⅲ patients, with statistical significances (P < 0.05). The accuracy of ultrasound, serum TPO-Ab and Tg-Ab combined to evaluate the histopathological grade of HT patients were higher than those of single and pairwise combination. Conclusion HT patients have increased serum TPO-Ab and Tg-Ab levels and ultrasound sonographic changes, and both are closely related to the clinical course of HT. The ultrasound combined with serum TPO-Ab and Tg-Ab can improve the diagnostic accuracy of the clinical course of HT.