Supplementary Material for: Prognostic value of amniotic fluid viral load to predict adverse outcome in pregnancies complicated by congenital Cytomegalovirus infection: a multicenter study

Introduction: To report the prognostic value of Cytomegalovirus (CMV) viral load in the amniotic fluid (AF) in predicting the outcome of infected pregnancies. Methods: Multicenter retrospective study involving 11 Italian referral centers from 2012 to 2021. Inclusion criteria were fetuses with confirmed congenital CMV infection. The primary outcome was the prognostic value accuracy of CMV qPCR in AF in predicting the risk of additional anomalies detected either at follow-up ultrasound or fetal MRI. The secondary outcome was prediction of post-natal clinical symptoms related CMV infection. Multivariate logistic regression and area under the curve (AUC) analyses were used to analyze the data. Results: 104 fetuses were included. Associated anomalies detected at follow-up ultrasound or fetal MRI were detected in 14.4% of cases (15/104). Mean AF CMV viral load was significantly higher in fetuses with compared to those without additional anomalies at follow-up ultrasound or fetal MRI (3346634.27±402582.95 vs 761934±222513,2 p1310520 copies/ml, a sensitivity of 66.7% and a specificity of 84.3% a positive likelihood ratio of 4.24. Once excluding fetuses with anomalies at ultrasound or MRI, the diagnostic performance of qPCR in identifying fetuses with symptomatic infection after birth was low, with an AUC of 0.586, Conclusion: CMV viral load at second trimester amniocentesis has a moderate accuracy for the occurrence of CMV related anomalies in fetuses with congenital infection and normal ultrasound at the initial diagnosis. Conversely, prediction of symptomatic infection is low.