Objectives To assess the possibility that gastric cancer can be screened non-endoscopically by serum tests: pepsinogen (PG)Ⅰ, PGⅡ, gastrin-17 (G-17) and H. pylori-IgG antibodies.Methods The study was performed on an observational case-control trial, and 310 subjects were recruited. Each of them underwent endoscopy with biopsies before serum tests were performed. These patients were further divided into three groups based on endoscopic and histological findings: 141 gastric cancer group (40 early gastric cancer and 101 advanced gastric cancer), 77 normal group and 92 atrophic gastritis. Serum samples of PGⅠ and Ⅱ, G-17, H. pylori-IgG antibodies were analyzed by ELISA.Results PGⅠ and PGR values were significantly decreased in gastric cancer group(28.74±11.55 μg/L, 1.66±1.01) as compared with those in normal group (123.99±32.25 μg/L, 10.09±1.89) and atrophic gastritis group (58.63±25.35 μg/L,4.36±2.57), respectively (P0.01). By using receiver operating curves (ROC), the cut-off points of PGⅠ and PGR values were 57.15 μg/L (sensitivity 99.3%, specificity 84.5%) and 2.99 (sensitivity 92.5%, specificity 89.0%) for the best discrimination of gastric cancer. G-17 level was significantly increased in gastric cancer group (20.86±8.24 pmol/L) as compared with those in normal group (10.39±9.25 pmol/L) and atrophic gastritis group (8.59±6.08 pmol/L), respectively(P0.01). For the best discrimination of gastric cancer, the cut-off point of G-17 was 14.61 pmol/L (sensitivity 75.2%, specificity 71.3%). PGⅠ and PGR values were significantly lower in patients with advanced gastric cancer than those in patients with early gastric cancer, while there was no difference in G-17 level between them. The positivity rate of H. pylori-IgG antibodies was higher in atrophic gastritis and gastric cancer groups than that in normal group(P0.01).Conclusions Based on markedly decreased serum PGI and PGR and increased serum G-17, gastric cancer can be screened and diagnosed endoscopically and histologically.