Objective: To analyze the clinical value and predictive difference of serum Golgi protein 73 (GP73) and serum autophagy-related protein p62 levels in the short-term prognosis of patients with hepatitis B virus-related acute-on-chronic liver failure (ACLF). Methods: Clinical data of admitted cases to our hospital from October 2018 to April 2020 were retrospectively analyzed. Simultaneously, there were 32 cases with HBV-related ACLF in group A, 65 cases with hepatitis B virus-related cirrhosis in group B and C (Child-Pugh Class A, 34 cases as B group, and Child-Pugh B/C class, 31 cases as group C), and another 30 healthy subjects served as the control group (group D). The serum GP73 and p62 levels of the four selected groups were measured. ACLF group patients were followed up for 3 months to analyze the prognosis of the patients. The serum GP73 and p62 levels of patients who died and survived during hospitalization were compared. The data were analyzed by one-way analysis of variance, independent sample t-test, and Pearson's correlation analysis. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of GP73 and p62 levels in surviving patients. Results: GP73 levels in the four groups A, B, C and D were (284.30 ± 70.55) ng/ml, (125.33 ± 20.57) ng/ml, (159.82 ± 31.20) ng/ml, and (45.46 ± 10.22) ng/ml, respectively. The p62 levels were (1.30 ± 0.35) ng/ml, (2.88 ± 0.58) ng/ml, (2.02 ± 0.545) ng/ml, and (4.68 ± 1.03) ng/ml, respectively. GP73 detection value was significantly higher in group A than the other three groups (P < 0.05). Group D had significantly lower value than the other three groups (P < 0.05), and group C had significantly higher value than group B (P < 0.05). The detection value of p62 in group A was significantly lower than the other three groups (P < 0.05). Group D had significantly higher value than the other three groups (P < 0.05), and group B had slightly higher value than group C, and the differences were statistically significant (P < 0.05). There was a negative correlation between GP73 and p62 (r = -0.695, P < 0.001). Survived patients GP73 level in the ACLF group was significantly lower than dead patients [(212.17 ± 22.47) ng/ml and (340.08 ± 32.91) ng/ml, t = 12.493, P < 0.05], and p62 level was significantly higher than dead patients [(1.46 ± 0.28) ng/ml and (1.18 ± 0.35) ng/ml, t = 2.445, P < 0.05]. According to the ROC curve analysis results, the area under the curve (AUC) of GP73 was 0.865, the AUC of p62 was 0.750, and the combined AUC of the both was 0.968. Conclusion: Both GP73 and p62 have a certain predictive value for the short-term prognosis of HBV-related ACLF patients, but the combination of the two indicators has a higher predictive value.