This study was carried out to determine the cutoff levels of serum pepsinogen (PG) I, II and their ratio of PG I/PG II for gastric cancer to establish a better screening system. Optimal cutoff levels for gastric cancer screening using serum pepsinogens were determined using Youden's index. The sensitivity, specificity and Youden's index for gastric cancer cases were calculated according to sex, age and the stage of gastric cancer, and the maximum Youden's index in each category was adopted as the cutoff level for gastric cancer screening using serum pepsinogens. The maximal Youden's index in all gastric cancer cases was 0.37, corresponding to a cutoff level of PG I < 40 (micrograms g/l) and PG I/PG II < 3.5. The sensitivity and specificity for gastric cancer cases of these cutoff levels were 0.50 and 0.87, respectively. In future, better criteria for gastric cancer screening have to be examined with the estimation of Youden's index in addition to other epidemiological methods such as ROC (receiver operating characteristic) curves and/or cost benefit analyses.