ObjectiveTo investigate the value of combined determination of hepatitis C virus (HCV) genotype, the alpha-fetoprotein variant AFP-L3, and P53 antibody in HCV-related hepatocellular carcinoma (HCV-HCC). MethodsA total of 84 patients with HCV-HCC who were diagnosed in our hospital from January 2016 to December 2019 were enrolled as HCV-HCC group, and 84 patients with benign liver diseases (hepatitis C and HCV liver cirrhosis) were enrolled as control group. The PCR-reverse dot blot hybridization technique was used to determine HCV genotype, ELISA was used to measure P53 antibody, and electrochemical luminescence was used to measure AFP-L3. The t-test and the Kruskal-Wallis H test were used for comparison between two groups; the chi-square test was used for comparison of categorical data between two groups. The logistic regression analysis and the receiver operating characteristic (ROC) curve were used to compare the value of each index in the diagnosis of HCV-HCC. ResultsCompared with the control group, the HCV-HCC group had a significantly higher proportion of patients with HCV 1b genotype or AFP-L3 and a significantly higher level of P53 antibody (χ2=5714, Z=-9.27, Z=-9.92, all P＜0.05). The logistic regression analysis showed that HCV genotype, AFP-L3, and P53 antibody had significant effects on HCV-HCC (all P＜0.05). The above indices were fitted to establish a model of Logit(Y)=-3.881+0031XAFP-L3(%)+0.043XP53+1218XHCV genotype, in which Y was the positive probability value of combined determination. In the screening of HCV-HCC, Y had a significantly larger area under the ROC curve than HCV genotype (0.945 vs 0.758, Z=6.17, P＜0001), AFP-L3 (0.945 vs 0.863, Z=3.97, P＜0.001), and P53 antibody (0.945 vs 0.887, Z=3.07, P=0.002). Y had higher AUC (0.945), sensitivity (90.90%), specificity (94.00%), positive predictive value (93.80%), negative predictive value (9116%), and diagnostic accuracy (92.44%) than each index alone. ConclusionHCV 1b genotype, AFP-L3, and P53 antibody level are associated with the risk of HCV-HCC, and the combined determination of the three indices has important clinical significance in the early diagnosis of HCV-HCC.