Background: A common and bothersome condition with a complicated pathophysiology, including neuropathy, is diabetes-related pruritus. The growth factor known as "brain-derived neurotrophic factor" (BDNF) affects how nerve cells survive, differentiate, and are maintained. Objectives: To measure the serum levels of BDNF and vitamin B12 (vitB12) in diabetic patients with generalized Pruritus. Patients and Methods: One hundred fifty participants were included in this case-control research. They were split into three equal groups: diabetes patients with chronic generalized pruritus (n=50), diabetic patients with no pruritus (n=50), and normal age-sex matched control subjects (n=50). Complete clinical information, including the duration of diabetes, the duration of pruritus, and a history of drugs, was gathered. Using the 12-Item Pruritus Severity Scale (12-PSS), the severity of the pruritus was evaluated. A thorough general and dermatological evaluation was performed. Lab tests included FBS, PPBS, and HbAIC. Additionally, ELISA was used to measure the serum levels of BDNF and vitB12. Results: Serum vitB12 and BDNF levels were significantly different among the study groups (P=0.000, P=0.000, respectively) with higher levels in diabetics compared to controls. Diabetics with pruritus exhibited higher vitB12, BDNF levels than diabetic non-pruritic group (P1=0.005, P1=0.000, respectively). Both vitB12 and BDNF showed significant positive correlation with 12-PSS (r=0.499, P=0.000; r=0.513, P=0.000, respectively). BDNF was positively correlated with vitB12 (r=0.518, P=0.000). There was a significant positive correlation between BDNF and the duration of diabetes and FBS, PPBS, HbA1C (P=0.000). Conclusion: Increased serum levels vitB12 and BDNF levels might explain the cause of chronic generalized pruritus in diabetic patients.