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To explore that if mid-regional sequence of pro-A-type natriuretic peptide (MR-proANP) may have a good value of diagnosis in heart failure with preserved ejection fraction (HFpEF) compared with N-terminal pro-B-type natriuretic peptide (NT-proBNP).Participants with cardiovascular disease who were enrolled in this study were divided into the nonheart failure (non-HF) group (n = 75), HFpEF group (n = 65), and HF with reduced ejection fraction (HFrEF) group (n = 50). The MR-proANP and NT-proBNP levels in plasma from all patients were measured by enzyme-linked immunosorbent assay.The plasma levels of MR-proANP and NT-proBNP in HFpEF and HFrEF groups were higher than those in non-HF group (P < .05). MR-proANP levels were significantly different (P < .05) in different New York Heart Association class patients with HFpEF. In the diagnostic analysis area under the curve of MR-proANP (0.844) was higher than that of NT-proBNP (0.518, P < .001). The left atrial volume index in the HFrEF group was higher than HFpEF group (P < .05); however, both of these groups had a higher index than non-HF group (P < .05).Results indicated that MR-proANP may be more sensitive and specific than NT-proBNP in diagnosing HFpEF. It may be used as a potential diagnostic biomarker in patients with HFpEF.