〔Abstract〕 Objective To analyze the application value of gastric serological biopsy in the diagnosis and treatment of gastric ulcer. Methods A total of 50 newly diagnosed patients with stomach discomfort admitted to Guangzhou Panyu Central Hospital from November 2020 to December 2021 were selected, who were pathologically confirmed as gastric ulcer by gastroscopic biopsy, and another 100 physical examination healthy people were selected as the physical examination group. The serum pepsinogen (PG)Ⅰ, PGⅡ and gastrin 17 (G-17) were detected by chemiluminescence method, and the ratio of PGⅠ/PGⅡ (PGR) was calculated, and the detection results of the two groups were compared. Results The levels of PGⅠ, PGⅡ, PGR and G-17 in the gastric ulcer group were significantly higher than those in the physical examination group (P < 0.05). The results of receiver operating characteristic curve (ROC) analysis showed that the area under curve (AUC) of serum PGⅠ, PGⅡ, PGR, G-17 in the diagnosis of gastric ulcer were 0.898, 0.807, 0.724, 0.856, respectively. The optimal critical values were 191.6 μg·L-1, 16.25 μg·L-1, 11.21, 6.70 pmol·L-1, respectively. The sensitivity and specificity of the combined detection of PGⅠ, PGⅡ, PGR and G-17 were 86.0% and 97.0%, respectively. Conclusion The increased levels of PGⅠ, PG Ⅱ, PGR and G-17 indicate an increased risk of gastric ulcer.