Background & Objective: A simple blood test (urea and creatinine) and a urine test may indicate renal function deterioration and presence of microalbuminuria, which is also the first clinical signs of renal dysfunction in patients with diabetes mellitus. This cost effective test easy to perform even in the absence of advance facilities in a hospital. That is why we planned to conducted this study to assess the comparison between of spot urine protein:creatinine ratio with 24-hour urinary protein in patients with type 2 diabetes mellitus. Materials and Methods: This prospective hospital based clinical study was conducted in the Department of General Medicine, Liaquat University of Medical & Health Sciences, Jamshoro, over a period of six months from 10th May 2018 to 9th November 2019 through a consecutive sampling technique. All the patients having age more than 18 years of both gender, and type 2 diabetes mellitus were enrolled in this study. Three cc blood was taken to determine the serum creatinine levels and twenty-four hour 2ml urine sample was also collected to determine the urinary protein levels. The proteinuria ≥300mg/dl in 24-hour urine sample was considered as significant proteinuria. Kappa statistics was used to find agreement between spot urine protein and 24 hours urinary protein. Results: A total 95 patients were evaluated and their mean age was 41.91±14.29 years, with male predominance (n = 66, 69.4%). Average 24 hour urinary protein was 1216.99±949.51mg and spot-urine evaluation of protein was 1919.12±2129.25mg. The agreement between spot urinary protein creatinine ratio and 24 hour urinary protein was found in 82.1% of cases through Kappa statistics and the calculated agreement between the two procedures was 0.975 which provides sufficient agreement to use spot urine protein:creatinine ratio in routine diagnosis of proteinuria. Conclusion: The study have shown that the protein:creatinine ratio for a random urine sample might be used to rule out the presence of significant proteinuria as defined by a quantitative measure of the 24-hour urine protein excretion.