Objective To explore the serum level of pepsinogen (PG) in health check-up and their distribution as well as relative factors. Methods Chemiluminescence was used to detect the serum level of PGⅠ、PGⅡ and the ratio of PGⅠ/Ⅱ (PGR) for 2993 subjects of health check-up. The latex test was employed to detect the serum Helicobacter Pylori (HP) antibody positive rate. Results The results showed that the serum PGⅠand PGR level of males was significantly higher than that of females. There was no significantly difference in males and females for the positive rate of PG detection. The PGⅠ and PGⅡ level increased with age. While the PGR showed a downward trend until age of 60 and then maintained at plateau. For the HP-positive group, the serum levels of PGⅠ and PGⅡ were significantly higher than those of the HP-negative group, while the PGR was significantly lower. Besides, the PG-positive rate gradually increased with age, while the HP-positive rate did not change significantly. Conclusion The serum PG levels are significantly skewed from normal distribution in the residents of Suzhou county, and affected by age and gender, as well as associated with HP infection. The PG positive rate is increased with age.