Chronic heart failure (CHF) is the leading cause of death worldwide. The regulatory interactions of long non-coding RNA (lncRNAs) and microRNAs (miRs) have important roles in multiple diseases. However, the clinical significance of the nuclear-enriched abundant transcript 1 (NEAT1)/miR-129-5p axis in CHF has remained elusive. The present study explored whether the NEAT1/miR-129-5p axis may be a suitable diagnostic and prognostic marker for CHF. The expression of lncRNA NEAT1 and miR-129-5p in the serum of patients with CHF was analyzed by reverse transcription-quantitative PCR. Furthermore, inter-indicator correlations were assessed by Pearson correlation coefficient analysis. Receiver operating characteristic (ROC) curves were generated to evaluate the ability of NEAT1, miR-129-5p and brain natriuretic peptide (BNP) to identify patients with CHF. The prognostic value of the NEAT1/miR-129-5p axis was analyzed by drawing Kaplan-Meier survival curves and by Cox logistic regression analysis. Baseline data were not significantly different between CHF (n=70) and control subjects (n=62). The serum level of NEAT1 was increased and the expression level of miR-129-5p was decreased in patients with CHF (all P<0.001). The ROC curves suggested that serum NEAT1 and miR-129-5p were of diagnostic value in patients with CHF and the combined diagnostic accuracy of NEAT1, miR-129-5p and BNP was significantly improved. Kaplan-Meier and multivariate Cox regression analysis suggested that low NEAT1 and high miR-129-5p were able to predict overall survival of patients with CHF (all P<0.01). In conclusion, the present study indicated that patients with CHF had increased NEAT1 and decreased miR-129-5p expression. The deregulated NEAT1/miR-129-5p axis may provide novel non-invasive biomarkers for the diagnosis and prognosis of CHF.