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Left ventricular (LV) volume is an important index for a wide range of cardiovascular diseases. In clinical assessment, echocardiography is one of the most frequently used imaging modalities. The aim of this study was to investigate the accuracy of the LV volume quantification through echo images based on a laboratory phantom with known volume. A tissue-mimicking phantom with the same ultrasound property as human blood and soft tissue was developed. It is considered to be a good reference for evaluating the accuracy of a semi-automated technique for LV volume measurement through 2D echo images. The volume measured by the semi-automated technique was compared with the theoretical volume and manual measured volume. The results showed a significant underestimation of the phantom volume when compared with the known volume: bias with known volume: -2.77ml (P=0.03); bias with manual measurement: -2.87ml (P=0.03).