Objective To study the level and application value of tumor marker (TM) in healthy population in Songjiang district of Shanghai. Methods A total of 37 695 healthy people from Songjiang District who have done physical examination in the physical examination center of Shanghai Songjiang District Central Hospital from January 2021 to December 2021 were included in this study. This study was a retrospective data analysis. Different TM detection levels and abnormal detection rates were analyzed, and the TM levels of people with different genders and different ages were compared. Results The results showed that the abnormal detection rate of cytokeratin 19 fragment (CY211) in all TM was the highest, and the abnormal rate of carcinoembryonic antigen (CEA) and CY211 in males was significantly higher than that in females (P<0.05), while the abnormal detection rate of carbohydrate antigen 199(CA199) in females was higher than that in males (P<0.05). There was no significant difference in the abnormal detection rate of alpha fetoprotein (AFP) in different gender groups (P>0.05). The abnormal detection rates of AFP, CEA, CA199 and CY211 were the highest in the age group over 61 years. The abnormal detection rates of total prostate specific antigen (TPSA), free prostate specific antigen (FPSA) and carbohydrate antigen 153(CA153) in the male population were also the highest in the age group over 61 years. The abnormal detection rates of CEA and male TPSA increased with age. The abnormal detection rate of carbohydrate antigen 125(CA125) in female population was the highest in the age group of 31~40. Conclusion There are significant differences in the levels of TM among different gender and age groups. Combined detection of multiple TM is beneficial to early detection and treatment of the disease.