Abstract Purpose Total kidney volume (TKV) is the most important imaging biomarker for quantifying the severity of autosomal-dominant polycystic kidney disease (ADPKD). 3D ultrasound (US) can accurately measure kidney volume compared to 2D US; however, manual segmentation is tedious and requires expert annotators. We investigated a deep learning-based approach for automated segmentation of TKV from 3D US in ADPKD patients. Method We used axially acquired 3D US-kidney images in 22 ADPKD patients where each patient and each kidney were scanned three times, resulting in 132 scans that were manually segmented. We trained a convolutional neural network to segment the whole kidney and measure TKV. All patients were subsequently imaged with MRI for measurement comparison. Results Our method automatically segmented polycystic kidneys in 3D US images obtaining an average Dice coefficient of 0.80 on the test dataset. The kidney volume measurement compared with linear regression coefficient and bias from human tracing were R² = 0.81, and − 4.42%, and between AI and reference standard were R² = 0.93, and − 4.12%, respectively. MRI and US measured kidney volumes had R² = 0.84 and a bias of 7.47%. Conclusion This is the first study applying deep learning to 3D US in ADPKD. Our method shows promising performance for auto-segmentation of kidneys using 3D US to measure TKV, close to human tracing and MRI measurement. This imaging and analysis method may be useful in a number of settings, including pediatric imaging, clinical studies, and longitudinal tracking of patient disease progression. Graphical abstract