Arteriovenous fistula (AVF), the preferred vascular access for hemodialysis, is associated with high failure rate. The aim of this study was to investigate the potential of AVF sound auscultation in providing quantitative information on AVF hemodynamic conditions. This single-center prospective study involved six patients with native radio-cephalic AVFs who underwent multiple follow-up visits. Doppler Ultrasound blood flow volume (BFV) assessment and electronic stethoscope-based sound recordings were performed during each visit, whereas MRIs were acquired 3days, 3weeks and 1year after surgery. Computational fluid dynamic (CFD) simulations were performed on patient-specific MRI-derived geometrical models. Higher values of median peak amplitudes ratios (high-low peak ratio-HLPR) were found to be associated with complex blood flow and velocity streamlines recirculation at systolic peak, and corresponding extended regions of high oscillatory shear index (OSI). On the contrary, lower values of HLPR were associated with laminar flow pattern and low values of OSI. Significant differences were observed in HLPR between subgroups with extended or limited areas with OSI > 0.1 (0.67 vs 0.31, respectively). Significant relationships were found between AVF sound intensity and brachial BFV (slope = 0.103, p < 0.01) as well as between longitudinal changes in brachial BFV and HLPR (slope = - 0.001, p < 0.01). Our results show that AVF sound can be exploited to extract fundamental information on AVF hemodynamic conditions, providing indication of the presence of complex hemodynamic and adequate BFV to perform hemodialysis. Sound analysis has therefore the potential to improve clinical AVF surveillanceand to ameliorate outcome.