[Purpose] To explore the significance of combined detection of serum alpha-fetoprotein(AFP) and Alpha-fetoprotein heterogeneity (AFP-L3) in the diagnosis for liver cancer. [Methods] Automatic chemiluminescent analysis technique was used to detect the AFP and AFP-L3 level in 185 cases with liver cancer,225 cases with benign liver lesion and 200 healthy controls.[Results] AFP and AFP-L3 levels in liver cancer group and benign liver lesion group were significantly higher than those in control group (P0.05) ; AFP and AFP-L3 levels in liver cancer group were significantly higher than those in benign liver lesion group (P0.05) . The sensitivity and specificity of AFP for diagnosing liver cancer were 68.11% and 80.94% respectively, and those of AFP-L3 were 83.78% and 96.00% respectively. The sensitivity and specificity of combined AFP with AFP-L3 was 93.51% and 78.35% . [Conclusion] Combined detection of AFP and AFP-L3 is positively significant for early diagnosis and differential diagnosis in liver cancer.