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Accurate aortic annulus (AoA) sizing is crucial for transcatheter aortic valve implantation planning. Three-dimensional (3D) transesophageal echocardiography (TEE) is a viable alternative to the standard multidetector row computed tomography (MDCT) for such assessment, with few automatic software solutions available. The aim of this study was to present and evaluate a novel software tool for automatic AoA sizing by 3D TEE.One hundred one patients who underwent both preoperative MDCT and 3D TEE were retrospectively analyzed using the software. The automatic software measurements' accuracy was compared against values obtained using standard manual MDCT, as well as against those obtained using manual 3D TEE, and intraobserver, interobserver, and test-retest reproducibility was assessed. Because the software can be used as a fully automatic or as an interactive tool, both options were addressed and contrasted. The impact of these measures on the recommended prosthesis size was then evaluated to assess if the software's automated sizes were concordant with those obtained using an MDCT- or a TEE-based manual sizing strategy.The software showed very good agreement with manual values obtained using MDCT and 3D TEE, with the interactive approach having slightly narrower limits of agreement. The latter also had excellent intra- and interobserver variability. Both fully automatic and interactive analyses showed excellent test-retest reproducibility, with the first having a faster analysis time. Finally, either approach led to good sizing agreement against the true implanted sizes (>77%) and against MDCT-based sizes (>88%).Given the automated, reproducible, and fast nature of its analyses, the novel software tool presented here may potentially facilitate and thus increase the use of 3D TEE for preoperative transcatheter aortic valve implantation sizing.Accurate AoA sizing is crucial for preoperative TAVI planning.A novel software tool for automatic AoA sizing by 3D TEE is presented.The software measurements were validated against standard manual MDCT values.It showed very good to excellent agreement with MDCT, being highly reproducible.This software may potentially facilitate the use of 3D TEE for preoperative TAVI sizing.