In the present study, the predictive value of glycosylated hemoglobin (HbA1c), microalbuminuria (24 h mAlb) and serum cystatin C (Cys-C) levels on the outcome of pregnancy in patients with gestational diabetes mellitus (GDM) was investigated. Samples of 144 females with GDM and 117 normal pregnant females as controls were selected for retrospective analysis. The following parameters were compared between the two groups: Levels of HbA1c, Cys-C and 24 h mAlb, maternal pregnancy outcome and adverse pregnancy rate. The predictive value of elevated 24 h mAlb, HbA1c and Cys-C regarding an adverse pregnancy outcome was then determined. Cys-C, 24 h mAlb and HbA1c levels in the GDM group were significantly higher than those in the control group (P<0.001). The adverse pregnancy rate in the GDM group was significantly higher than that in the control group (40.97 vs. 16.24%; P<0.001). Logistic regression and receiver operating characteristics (ROC) analyses indicated that, in subjects with GDM, HbA1c, Cys-C and 24 h mAlb levels were closely associated with adverse pregnancy outcomes (P<0.050) and may be considered as predictors for an adverse pregnancy outcome (risk ratio >1). Linear correlation analyses indicated that HbA1c, Cys-C and 24 h mAlb were negatively correlated with the neonatal Apgar scores (r=-0.509,-0.954 and-0.954, respectively; P<0.001). According to ROC analysis, the combined predictive sensitivity of HbAlc, Cys-C and 24 h mAlb for adverse pregnancy outcome in patients with GDM was 96.49% and the specificity was 77.19%. The increase in HbAlc, Cys-C and 24 h mAlb levels is expected to be an effective predictor of adverse pregnancy outcomes in high-risk pregnant women.