Objective To study the detection value of serum alpha-L-fucosidase(AFU) and alpha fetoprotein 3(AFP-L3) in primary hepatocellular carcinoma(HCC). Methods 100 patients with primary HCC were enrolled as HCC group, 54 cases without HCC were selected as non HCC group, 100 healthy persons were selected as healthy group.Serum AFU and AFP-L3 levels in three groups were determined.The serum AFU was measured by the chemical colorimetry, and the AFP-L3 level was determined by chemiluminescence immunoassay. Results The serum levels of AFU in HCC group, non HCC group and healthy group were (71.61±3.01)U/L, (11.06±2.15)U/L, (8.54±1.28)U/L.The serum levels of AFP-L3 in HCC group, non HCC group and healthy group were (30.14±2.93)%, (3.27±0.83)%, (2.77±0.62)%.The serum levels of AFU and AFP-L3 in HCC group were significantly higher than those in non HCC group and healthy group (t=13.172, 9.770, 16.036, 7.160, all P 0.05). The positive rates of AFU were 87.00%, 9.26% and 3.00% in HCC group, non HCC group and healthy group.The positive rate of AFU in HCC group was significantly higher than that in non HCC group and healthy group (χ2=34.06, 57.62, all P 0.05). The positive rates of AFP-L3 were 89.00%, 7.41% and 2.00% in HCC group, non HCC group and healthy group.The positive rate of AFP-L3 in HCC group was significantly higher than that in non HCC group and healthy group (χ2=41.06, 77.62, all P 0.05). There were no significant differences in serum tumor diameter about AFU, AFP-L3(t=0.167, 0.579, 0.201, 0.601, all P>0.05), and there were significant differences about TNM stage, lymph node metastasis, and tumor thrombus (t=6.156, 9.105, 10.161, 8.120, 9.770, 12.821, all P<0.05). Conclusion Serum AFU and AFP-L3 can not only help the diagnosis of primary liver cancer, but also can determine the prognosis of patients. Key words: Liver neoplasms; Alpha-L-fucosidase; Alpha fetoprotein 3