[Purpose] This study compared the differences in electrophysiological characteristics of normal muscles versus muscles with latent or active myofascial trigger points, and identified the neuromuscular physiological characteristics of muscles with active myofascial trigger points, thereby providing a quantitative evaluation of myofascial pain syndrome and clinical foundational data for its diagnosis. [Subjects] Ninety adults in their 20s participated in this study. Subjects were equally divided into three groups: the active myofascial trigger point group, the latent myofascial trigger point group, and the control group. [Methods] Maximum voluntary isometric contraction (MVIC), endurance, median frequency (MDF), and muscle fatigue index were measured in all subjects. [Results] No significant differences in MVIC or endurance were revealed among the three groups. However, the active trigger point group had significantly different MDF and muscle fatigue index compared with the control group. [Conclusion] Given that muscles with active myofascial trigger points had an increased MDF and suffered muscle fatigue more easily, increased recruitment of motor unit action potential of type II fibers was evident. Therefore, electrophysiological analysis of these myofascial trigger points can be applied to evaluate the effect of physical therapy and provide a quantitative diagnosis of myofascial pain syndrome.