Objective: To investigate the value of serum matrix metalloproteinase-2 (MMP-2), alpha fetoprotein (AFP) and model for end-stage liver disease (MELD) score in prognosis assessment of patients with HBV-related acute on chronic liver failure (HBV-ACLF). Methods: Clinical data of 90 HBV-ACLF patients were collected. The patients were divided into a survival group and a death group according to the clinical outcome. The differences of serum MMP-2 and AFP levels and MELD scores between the 2 groups were compared. Logistic regression analysis of the relationship between the indicators and the prognosis of patients and the prognostic value of MMP-2, AFP level and MELD score in patients with HBV-ACLF were analyzed. Results: The serum MMP-2 level and MELD score of death group were higher than those in the survival group, while the serum AFP level was significantly lower than the survival group (P<0.05). Serum MMP-2 level, AFP level and MELD score were important factors affecting the prognosis of HBV-ACLF patients (P<0.05). Serum AFP level in HBV-ACLF patients was significantly negatively correlated with MMP-2 level and MELD score, while MMP-2 level was positively correlated with MELD score (P<0.05). The area under the ROC curve of MMP-2, AFP level and MELD score predicting the short-term prognosis of HBV-ACLF patients was 0.832, 0.846 and 0.858, respectively. The predictive sensitivity of combined test of MMP-2, AFP level and MELD score was significantly higher than single test (P<0.05), while the specificity of combined test was not statistically different from single test (P>0.05). Conclusion: Serum MMP-2, AFP level and MELD score are important risk factors affecting the prognosis of HBV-ACLF patients. The detection of these indicators is helpful to evaluate the prognosis of patients.