Clinical Potential of HCMV-Encoded Serum miRNAs in Predicting Intrahepatic Cholestasis of Pregnancy: A Retrospective Analysis.

Intrahepatic cholestasis of pregnancy (ICP) is a prevalent liver disorder that typically occurs during the second and third trimesters of pregnancy and is associated with adverse perinatal outcomes. Currently, total bile acid (TBA) serves as the primary diagnostic biomarker in clinical settings. but its sensitivity and specificity are limited. The relationship between microRNA (miRNA) encoded by human cytomegalovirus (HCMV) and ICP remains unclear. This study retrospectively analyzed serum samples from 151 pregnant women diagnosed with ICP and 158 age-matched healthy controls between October 2021 and July 2024. A three-phase research design (training set, validation set, and testing set) was implemented to comprehensively assess the expression levels of 24 HCMV-encoded miRNAs using TaqMan probe-based RT-qPCR, a highly specific detection method. The receiver operating characteristic (ROC) curve and logistic regression analysis were used for evaluation. Meanwhile, serum HCMV IgG/IgM levels and whole-blood HCMV DNA copy numbers were measured, and potential target genes were predicted. RT-qPCR results revealed six miRNAs (hcmv-miR-UL22A-5p, hcmv-miR-US4-5p, hcmv-miR-US4-3p, hcmv-miR-UL112-5p, hcmv-miR-UL148D and hcmv-miR-US25-2-5p) were significantly upregulated in ICP patients (p < 0.001) and their combined detection yielded the maximum area under the curve(AUC) of 0.814. Logistic regression analysis indicated that they could potentially serve as independent risk factors for ICP. Serological analysis showed comparable HCMV IgG positivity rates between the two groups (96.8% in controls vs. 97.4% in ICP cases). Although HCMV DNA was undetectable in both groups, the IgM level in ICP pregnant women was increased (p < 0.05). Target gene prediction suggests that these miRNAs may be involved in the pathogenesis of ICP by regulating multiple host genes. The panel of six HCMV miRNAs identified in this study holds promise as a novel liquid biopsy-based diagnostic molecular marker, which could enhance the accuracy of ICP diagnosis in clinical practice and provide molecular evidence supporting the role of latent viral infection in ICP development.