Objective To explore the value of the urine N-acetyl-beta-D-glucosaminidase and β2- microglobumin in diagnosis of patients with stage 2-4 chronic kidney disease. Methods Urine N-acetyl-beta-D-glucosaminidase and β2- microglobumin in 144 patients with chronic kidney disease and 50 healthy controls were analyzed. The eGFR was calculated using the modified Modification of Diet in Renal Disease (MDRD) equation. Results There were statistical significance between the patients with chronic kidney disease group and control group with urine N-acetyl-beta-D-glucosaminidase and β2- microglobumin. The level of the urine N-acetyl-beta-D-glucosaminidase and β2- microglobumin were increased in the chronic kidney disease stage 2-4. The urine N-acetyl-beta-D-glucosaminidase levels were 35.3±12.4 mg/dL in stage 3a and 45.4±11.5 mg/dL in stage 3b. The urine β2- microglobumin levels were 0.45±0.35 mg/dL in stage 3a and 1.20±0.88 mg/dL in stage 3b. On the Pearson’s analysis, the urine N-acetyl-beta-D-glucosaminidase and β2- microglobumin were found to be positively correlated to creatinine and negatively correlated to eGFR. Conclusion The urine N-acetyl-beta-D-glucosaminidase and β2- microglobumin might be useful laboratory markers to assess the renal damage of the chronic kidney disease.