Persistent respiratory syncytial virus (RSV) infections have been associated with the exacerbation of chronic inflammatory diseases, including chronic obstructive pulmonary disease (COPD). This virus infects the respiratory epithelium, leading to chronic inflammation, and induces the release of mucins and the loss of cilia activity, two factors that determine mucus clearance and the increase in sputum volume. In this study, an automatic method has been established to determine the ciliary motility activity from cell cultures by means of optical flow computation, and has been applied to 136 control cultures and to 144 RSV-infected cultures. The control group presented an average of cell surface with cilia motility per field of 41 ± 15 % (mean ± standard deviation), while the infected group presented a 11 ± 5 %, t-Student p<0.001. The cutoff value to classify a infected specimen was <17.89 % (sensitivity 0.94, specificity 0.93). This methodology has proved to be a robust technique to evaluate cilia motility in cell cultures.