The aim of this study is to determine whether stereotactic body radiotherapy for multiple vertebral metastases treated with a single isocenter results in greater intrafraction errors than stereotactic body radiotherapy for single vertebral metastases and to determine whether the currently used spinal cord planning organ at risk volume and planning target volume margins are appropriate. Intrafraction errors were assessed for 65 stereotactic body radiotherapy treatments for vertebral metastases. Cone beam computed tomography images were acquired before, during, and after treatment for each fraction. Residual translational and rotational errors in patient positioning were recorded and planning organ at risk volume and planning target volume margins were calculated in each direction using this information. The mean translational residual errors were smaller for single (0.4 (0.4) mm) than for multiple vertebral metastases (0.5 (0.7) mm; P = .0019). The mean rotational residual errors were similar for single (0.3° (0.3°) and multiple vertebral metastases (0.3° (0.3°); P = .862). The maximum calculated planning organ at risk volume margin in any direction was 0.83 mm for single and 1.22 for multiple vertebral metastases. The maximum calculated planning target volume margin in any direction was 1.4 mm for single and 1.9 mm for multiple vertebral metastases. Intrafraction errors were small for both single and multiple vertebral metastases, indicating that our strategy for patient immobilization and repositioning is robust. Calculated planning organ at risk volume and planning target volume margins were smaller than our clinically employed margins, indicating that our clinical margins are appropriate.