[show abstract][hide abstract] ABSTRACT: The purpose of this study is to evaluate the 3-dimensional target localization accuracy of cone-beam computed tomography (CBCT) using an on-board imager (OBI). An anthropomorphic pelvis phantom was used to simulate a range of offsets in the three translational directions and rotations around each of the three axes. After a translational or rotational offset was applied, a CBCT scan of the phantom was followed by image registration to detect the offsets in six degrees. The detected offsets were compared to the offset actually applied to give the detection error of the phantom position. Afterwards, the phantom was positioned by automatically moving the couch based on the detected offsets. A second CBCT scan followed by image registration was performed to give the residual error of the phantom positioning. On the average the detection errors and their standard deviations along the lateral, longitudinal and vertical axis are 0.3 ± 0.1, 0.3 ± 0.1 and 0.4 ± 0.1 mm respectively with respect to translational shifts ranging from 0 to 10 mm. The corresponding residual errors after positioning are 0.3 ± 0.1, 0.5 ± 0.1 and 0.3 ± 0.1 mm. For simulated rotational shifts ranging from 0 to 5 degrees, the average detection error and their standard deviation around lateral, longitudinal, and vertical axes are 0.1 ± 0.0, 0.2 ± 0.0, and 0.2 ± 0.0 degrees respectively. The residual errors after positioning are 0.4 ± 0.1, 0.6 ± 0.1, and 0.3 ± 0.1 mm along the lateral, longitudinal and vertical directions. These results indicate that target localization based on CBCT is capable of achieving sub-millimeter accuracy.