Objective To evaluate the clinical application of combined detection of golgi glycoprotein 73（GP-73），percentage of AFP heterogeneity（AFP-L3 %）and total bile acid（TBA）in diagnosis of primary hepatocellular carcinoma. Methods 100 cases of primary hepatocellular carcinoma patients（primary hepatocellular group）, 50 cases of patients with cirrhosis（cirrhosis group） and 50 normal controls（control group）were involved in this study. The concentration of GP-73 and AFP-L3 were measured by enzyme-linked immunosorbent assay and the level of alpha-fetoprotein（AFP）was detected by radioimmunoassay and the level of TBA was measured by enzymatic cycle. Results The level of GP-73，AFP-L3% and TBA in primary hepatocellular carcinoma group were significantly higher than that in the cirrhotic group and control group（P <0.05）. The positive rate of three tumor markers combined determination was 95.0%，which was significantly higher than that of individual determination of GP-73、AFP-L3 and TBA（P <0.05）. The positive rate of individual determination of GP-73, AFP and TBA was obviously higher than that of AFP（P <0.05）. Conclusion The combined detection of serum GP-73, AFP-L3% and TBA could improve the positive rate of primary hepatocellular carcinoma and has important clinical significance in early diagnosis of primary hepatocellular carcinoma.