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Background: One of the most common metabolic diseases, Type 2 Diabetes Mellitus (T2DM), is caused by a combination of two basic factors: insufficient insulin secretion by pancreatic -cells or a failure of insulin-sensitive tissues to respond adequately to insulin. The aim of this paper was to assess the diagnostic accuracy of serum nesfatin-1 in type 2 diabetes mellitus (T2DM). Methods: Sixty patients with T2DM were recruited from (Baquba Teaching Hospital) in Iraq during the period (from November 2021 - March 2022). T2DM group was classified into 30 newly diagnosis patients (without treatment) and 30 ongoing diabetic patients (with treatment) for comparison, as well 30 healthy individuals were included as a control. The ELISA Kit was used to measure serum nesfatin-1 and serum insulin, fasting serum glucose, and lipid profile test were measured through spectrophotometric, instead of HbA1c was determined using HPLC method. Results: The concentration of serum nesfatin-1 in the T2DM group was significantly lower than that of the healthy subjects (p < 0.05). There was a significant difference in the serum nestafin-1 concentrations between newly diagnosis and ongoing T2DM patients. There were substantial negative connections between serum Nesfatin-1 concentration and HOMA-IR, as well as strong negative correlations between serum nesfatin-1 and serum insulin level. The concentration of serum Nesfatin-1, on the other hand, had no significant association with the anthropometries measurements and biochemical parameters. The area under the curve was excellent (AUC = 0.827, p = 0.0001), with high diagnostic accuracy (86.2) in differentiating newly diagnosis T2DM from the healthy subject group. Conclusion: Nesfatin-1 levels in the sera of diabetic patients was shown to be lower and nesfatin-1 levels were shown to be significantly linked to newly diagnosed patients. Highlight: • Nesfatin-1 is a novel biomarker candidate in type 2 diabetes mellitus. • Nesfain-1 can be used to differentiate T2DM from healthy subject. • The nesfatin-1 change depends on the severity of diseases (disease severity depended on increased HOMA-IR) of T2DM. [ABSTRACT FROM AUTHOR]