Abstract: Objective：To establish the indirect reference intervals for serum cytokeratin 19 fragment(Cyfra21-1), squamous cell carcinoma antigen(SCCA) and carcinoembryonic antigen(CEA) from hospital-stored data. Methods：All the test results of serum Cyfra21-1, SCCA and CEA stored in the laboratory information system(LIS) of the Affiliated Hospital of Nantong University during April 2014 to March 2015 were collected, screened and linearly fitted to establish the indirect reference intervals. Meanwhile, a total of 454 subjects who performed medical examination in the Affiliated Hospital of Nantong University during January 2015 to March 2015 were enrolled, and the serum Cyfra21-1, SCCA and CEA reference intervals were established with non-parametric methodology recommended by CLSI C28-A3. Results：The indirect reference intervals for serum Cyfra21-1, SCCA and CEA based on hospital-stored data were 0~2.87 ng/mL, 0~1.76 ng/mL and 0~3.77 ng/mL, while those based on the healthy subjects were 0~2.84 ng/mL, 0~1.38 ng/mL and 0~3.60 ng/mL. Conclusion：The establishment of indirect reference intervals based on hospital-stored data may save time and labor, which may be used to verify the current reference intervals in laboratories.