Diabetic neuropathy (DN) is a major complication associated with diabetes mellitus (DM) and results in fatigue. We investigated whether type 2 diabetic patients with or without neuropathy experienced muscle fatigue and determined the most influencing factor on muscle fatigue. Overall, 15 out of 25 patients with type 2 DM were diagnosed with DN using a nerve conduction study in the upper and lower extremities, and the composite score (CS) was calculated. We obtained the duration of DM and body mass index (BMI) from subjects, and they underwent a series of laboratory tests including HbA1c, fasting plasma glucose, triglycerides, and high- and low-density lipoprotein. To qualify muscle fatigue, this study used surface electromyography (sEMG). Anode and cathode electrodes were attached to the medial gastrocnemius. After 100% isometric maximal voluntary contracture of plantarflexion, the root mean square, median frequency (MDF), and mean power frequency (MNF) were obtained. We showed a correlation among laboratory results, duration of DM, BMI, CS, and parameters of muscle fatigue. The duration of DM was related to fatigue of the muscle and CS (p < 0.05). However, CS was not related to fatigue. The MDF and MNF of muscle parameters were positively correlated with HbA1c and fasting plasma glucose (p < 0.05). In conclusion, we suggest that the duration of DM and glycemic control play important roles in muscle fatigue in patients with DN. Additionally, sEMG is useful for diagnosing muscle fatigue in patients with DN.