SummaryBackgroundObjective gait analysis is becoming more popular as a tool assisting veterinarians during the clinical lameness exam. At present, there is only limited information on the effect of misplacement of markers/motion‐sensors.ObjectivesTo investigate and describe the effect of marker misplacement on commonly calculated pelvic symmetry parameters.Study designExperimental study.MethodsEach horse was equipped with custom‐made devices consisting of several reflective markers arranged in a predefined manner with a reference marker correctly positioned regarding the anatomical landmark and several misplaced markers along the sagittal and transverse planes. Linear regression analysis was used to estimate the effect of marker misplacement.ResultsFor the tubera sacrale, each cm of left/right misplacement led to a difference in minimum position of the pelvis (PDmin) of ±1.67 mm (95% CI 1.54–1.8 mm) (P<0.001); maximum position of the pelvis (PDmax) was affected by ±0.2 mm (95% CI 0.071–0.33 mm) (P = 0.003). With respect to cranial/caudal misplacement, each cm of misplacement resulted in a PDmin difference of ±0.04 mm (95% CI −0.09 to 0.16 mm) (P = 0.56) and a PDmax difference of ±0.008 mm (95% CI −0.13 to 0.12 mm) (P = 0.9). For the tubera coxae, each cm of vertical misplacement led to a difference in the displacement amplitude between left and right tubera coxae (Hip‐Hike_Diff) of ±1.56 mm (95% CI 1.35–1.77 mm) (P<0.001); for the cranial/caudal misplacement, this was ±0.82 mm (95% CI 0.66–0.97 mm) (P<0.001).Main limitationsOnly three horses were used in this experiment and the study design did not permit to determine the influence of marker misplacement on the evaluation of different degrees of lameness.ConclusionsMarker misplacement significantly affects calculated symmetry parameters of the pelvis. The observed errors are overall small but significant. In cases of mildly asymmetrical horses, this error might influence the decision‐making process whereas in more severe asymmetries, the influence of the error effect may become less significant.