This study aims to investigate the causes of abnormal riding comfort (excessive and W-shaped distribution of riding comfort) in dynamic tracking tests of low-floor trams and propose appropriate improvement measures. A multibody system dynamics model of the tram is established, and the accuracy of the model is verified through a dynamic tracking test. The results reveal that the primary cause of excessive comprehensive riding comfort is due to the large amplitude of the measured irregularity and the low equivalent conicity of the wheel-rail contact relationship. The main reason for the W-shaped distribution of riding comfort is the weak inter-vehicle constraints and the lack of lateral energy-absorbing devices. Based on these findings, the following measures are recommended to improve riding comfort: (1) reducing lateral stiffness of the secondary suspension, (2) modifying structural parameters of the M3, (3) modifying the joint type between TP2 and M3, (4) introducing a damper between the middle car. By taking these measures, the riding comfort of low-floor trams has been effectively improved.