Cancer is the second leading cause of death and gastric cancer is the fourth most common cancer type worldwide. Investigation of autoantibodies in cancer patients has been a popular research area in recent years. The aim of the current study was to investigate carbonic anhydrase I and II (CA I and II) autoantibodies in the plasma of subjects with gastric cancer based on the information and considerations of autoimmune relation of gastric cancer. Anti-CA I and II antibody levels were investigated by ELISA in plasma samples of fifty two patients with gastric cancer and thirty five healthy peers. Anti-CA I and II antibody titers of the gastric cancer group were significantly higher compared with the control group (p = 0.004, p = 0.0001, respectively). Plasma anti-CA I levels of the metastatic group were lower than the non-metastatic group and this difference was found statistically significant (p < 0.05), but there was no statistical difference between plasma anti-CA II levels of the groups. CA I and II autoantibody titers in patients with gastric cancer were found higher compared to healthy subjects and the results suggest that these autoantibodies may be involved in the pathogenesis of gastric cancer.