Objective To investigate the clinical value of serum brain natriuretic peptide (BNP), soluble growth stimulated expressed gene 2 (sST2), interleukin-6 (IL-6), and cardiac enzyme panel in the diagnosis of heart failure. Methods Clinical data of 100 patients with HF treated at The First People’s Hospital of Zhaoqing from January 2022 to December 2024 and 50 healthy individuals undergoing physical examination during the same period were retrospectively analyzed. Fasting venous blood samples were collected from HF patients on the morning of the day after admission and from healthy controls on the morning of physical examination. Serum BNP, sST2, IL-6, and cardiac enzyme panel levels were measured. Pearson or Spearman correlation analysis was used to assess the associations of these markers with left ventricular ejection fraction (LVEF) and New York Heart Association (NYHA) functional class. Receiver operator characteristic (ROC) curves were used to evaluate diagnostic performance, and a combined Logistic regression model was established. Results Serum levels of BNP, sST2, IL-6, and cardiac enzyme panel markers in the heart failure group were significantly higher than those in the control group (all P＜0.05). BNP, sST2, and IL-6 were positively correlated with each other, negatively correlated with LVEF, and positively correlated with NYHA functional class (all P＜0.001). Among the single indicators, BNP showed the best diagnostic performance, with an area under the curve (AUC) of 0.902. The combined model of BNP, sST2, and IL-6 yielded an AUC of 0.948, with a sensitivity of 92.0% and a specificity of 84.0%, outperforming any single indicator. Conclusion Serum BNP, sST2, IL-6 combined with cardiac enzyme panel has high diagnostic value for HF. Multi-marker combination improves diagnostic accuracy and may aid early identification and risk stratification.