Objective. Volumetric flow measurement with Doppler ultrasound is useful in assessing blood flow as part of an evaluation of arteriovenous fistula maturity in patients undergoing hemodialysis. In this study, we assessed both accuracy and variability in volumetric flow measurements obtained using modern and commercially available ultrasound systems and an in vitro experimental setup. Methods. Volumetric flow measurements using duplex ultrasound were obtained by 3 users operating 5 different systems for randomized flow in the range of 100 to 1000 mL/min. Users performed 3 consecutive measurements at a given flow rate. Data were analyzed using statistical techniques to assess measurement accuracy and variability. Results. Over the span of flow rates studied, the root mean square error (RMSE) for the 5 ultrasound systems ranged from 38.8 to 79.7, 36.8 to 52.0, 73.0 to 85.3, 26.7 to 44.6, and 43.9 to 93.5 mL/min. Corresponding average RMSE values were 60.3, 42.7, 81.1, 37.2, and 64.4 mL/min, respectively. A linear regression analysis of mean interobserver measurements revealed an excellent correlation for all ultrasound systems (r2 > 99.1%). Assessment of intraobserver measurements revealed no statistically significant differences for any ultrasound system evaluated (P > .94). Comparison of interobserver measurements indicates no statistically significant differences between any of the 5 systems (P > .14). Conclusions. Modern ultrasound systems are reasonably accurate in blood flow measurement in an experimental setup mimicking clinically relevant blood flow ranges in a hemodialysis fistula. Users need adequate training and experience to perform multiple measurements and use appropriate techniques to minimize errors in flow measurement. © 2009 by the American Institute of Ultrasound in Medicine.