Abstract: Objective To investigate the clinical application value of blood glucose (Glu), glycosylated hemoglobin (HbA1c), and glycosylated serum protein (GSP) three combined detection. Methods Eighty patients clinically diagnosed as having type 2 diabetes mellitus underwent Glu, HbA1c and GSP determinations before and after treatment, and another 80 healthy subjects were recruited as the control group. The results were analyzed statistically. Results In diabetic patients the fasting Glu, HbA1c and GSP levels were significantly higher than those in normal control group. And in diabetic group the fasting blood glucose and HbA1c and GSP were positively correlated; the results of the three determinations were statistically significantly different (P<0.05). Conclusions Glu, HbA1c and GSP three combined detection is important in early diagnosis of type II diabetes mellitus; it helps assess therapeutic results, control and reduce the incidence and development of diabetic complications.