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Aortic stenosis (AS) is the most common native valve disease. The assessment of its severity is routinely performed using transthoracic 2D echocardiography. In particular, aortic valve area (AVA) is usually assessed by continuity equation, although this measurement relies on geometrical assumptions, or by planimetry on transesophageal echocardiography. Accordingly, the aim of this study was to develop and test a new technique to assess the severity of AS using data acquired with real-time 3D matrix transesophageal echocardiographic (MTEE) probe. We studied 20 patients undergoing clinically indicated TEE. MTEE imaging was performed in 10 subjects with normal mitral and normal aortic valves and in 10 patients with moderate to severe AS (AVA < 1.4 cm2) and normal mitral valve. Our technique allowed quantitative volumetric measurements of dynamic AV parameters, and thus constitutes a new tool for objective assessment of the severity of AS.