Abstract: Objective To establish a quantitative chemiluminescence immunoassay for glycocholic acid. Methods The competition model and the microplate coating technology were used to assure the test kits have same clinical diagnostic value by comparing the clinical result with the reference kits. Results The linear range of the test kit was 0.25 to 40μg/ml. The sensitivity was 0.05μg/ml. The coefficients of variation for intra-assay were 7.1% and 7.4%, and for inter-assay were 11.3% and 15.74% respectively. The recovery rate of the samples was in the range of 85% to 115%. The cross-reactivity rate of hyaluronic acid, IVC, PⅢNP, Laminin, Cholic acid and Chenodeoxycholic with Glycocholic acid was 0.010%～1.480%. The results determined with test kits were consistent with RIA kits, and the correlation coefficient r was 0.947 between the two groups of detection results. Conclusion The test chemiluminescence immunoassay kit could provide reliable indicators for clinical diagnosis and research for chronic hepatitis, cirrhosis, liver cancer and digestive disease. Meanwhile, combined test of glycocholic acid with the CIV, HA and PA could have very important clinical significance for the diagnosis, treatment and prognosis of cirrhosis.