The primary component of ultra-precision machining machines and measuring instruments is a high-precision spindle, and spindle error affects the accuracy of these machines. Traditional spindle error measurement methods often require a standard device as a reference, which inevitably reduces the accuracy of spindle error measurement, while the error separation technique is tedious and time-consuming. This study proposed a spindle error measurement method based on a composite laser target, which can measure the motion error of the spindle, including axial, radial, and tilt error in terms of five degrees of freedom. In this method, a composite laser target containing a laser point source and laser collimated beam was constructed and mounted on the spindle as a reference frame to mark the position of the spindle during rotation. Consequently, the error of the spindle was obtained via measurements of the position and angle of the composite laser target. Differential confocal microscopy technique was employed for measuring the axial position of the laser point source to obtain the axial error of the spindle, whereas the radial position was measured using a conventional microscope optical path to obtain the radial error of the spindle. Further, the angle of laser collimation was measured using a collimation measurement optical path to obtain the tilt error of the spindle. The measurement resolution of the proposed method for spindle error was verified through suitable experiments, yielding resolutions of 4 and 2 nm for axial and radial errors, respectively, and 0.2 μrad in case of tilt error. In addition, the method was tested on an air spindle to demonstrate the feasibility of using the method on machines. Thus, it was demonstrated that the proposed method can be used to obtain the spindle error by optical standard devices without additional error separation techniques, particularly for a spindle in ultra-high precision measurement equipment. Further, it facilitates real time monitoring or evaluation and calibration of the spindle error. This method is expected to serve several important practical applications in the field of ultra-precision machining and measurement. • Spindle error measurement method based on a composite laser target proposed. • Method eliminates reliance of accuracy on standard device used. • Motion error of spindle: axial, radial, tilt in terms of five degrees of freedom. • Resolution of proposed method verified through suitable experiments. • Practical applications in the field of ultra-precision machining and measurement.