Objective To study the diagnostic performances of serum anti-p53 antibody(anti-p53)detection as well as combined detection of serum anti-p53 with(AFP)for hepatocellular carcinoma(HCC). Methods Serum anti-p53 and AFP of 50 patients with HCC, 50 patients with liver benign lesion(LBL)and 50 healthy controls(HC)were detected by enzyme linked immunosorbent assay(ELISA)and electrochemiluminescence immunoassay(ECL), respectively. Results The level and positive rate of serum anti-p53 in HCC group were significantly higher than that in LBL group(P<0.05)and HC group(P<0.01). The sensitivity, specificity and area under ROC curve(AUC)of serum anti-p53 for HCC were 36.0%, 91.0% and 0.630(95%CI 0.547-0.707), respectively. The sensitivity, specificity and AUC of serum AFP for HCC were 54.0%, 83.0% and 0.685(95%CI 0.604-0.758), respectively. The sensitivity and AUC of combined detection of anti-p53 with AFP for HCC was improved to 76.0%(specificity 80.0%)and 0.770(95%CI 0.694-0.835), respectively. Conclusions Anti-p53 has an ideal diagnostic value for HCC. The combined detection of anti-p53 with AFP might contribute to the marked improvement of diagnostic performance for HCC, with the evidence of improved sensitivity and enough high specificity for HCC.