Objective To explore the application value of Apolipoprotein A1(APOA1) and cholinesterase(CHE) markers in the diagnosis and severity of intrahepatic cholestasis of pregnancy (ICP). Methods This retrospective case-control study was conducted with 100 ICP patients and 100 normal pregnant women. Protein molecular marker APOA1 and CHE were compared between the groups.Results APOA1 and CHE levels in the ICP group were significantly lower than those in the normal group(P＜0.05), the levels of two markers were negatively associated with that of total bile acids.The areas under the receiver operating characteristic (ROC) curves (AUCs) of APOA1 and CHE in diagnosis of ICP were 0.871 and 0.782, respectively, the areas under the receiver operating characteristic (ROC) curves (AUCs) of APOA1 and CHE in severity of ICP were 0.843 and 0.773, respectively. Multiple logistic regression showed that a combination of the levels of the two markers optimized the AUC (0.909), thus more reliably diagnosing ICP. Multiple logistic regression showed that a combination of the levels of the two markers optimized the AUC (0.891), thus more reliably to determine the severity of ICP.Conclusion Combined detection of APOA1 and CHE markers can be used as novel noninvasive biomarkers to evaluate the diagnosis and severity of ICP. The findings of this study can improve the diagnosis rate of ICP disease and effectively improve the perinatal outcome of mothers and infants, which has important theoretical significance and clinical application value.