To investigate whether invasive procedures performed in the presence of human cytomegalovirus (HCMV) DNA in maternal peripheral blood (HCMV DNAemia) represent a risk for iatrogenic transmission of HCMV infection to the fetus, 194 pregnant women undergoing prenatal diagnosis because of a primary HCMV infection and their 199 fetuses were investigated. Overall, 27 (37%) of 73 mothers of uninfected fetuses and 22 (37%) of 59 mothers of infected fetuses were HCMV DNAemia-positive at amniocentesis. Of the 8 mothers of the 8 fetuses with false-negative amniocentesis results, 4 were DNAemia-positive and 4 were DNAemia-negative at amniocentesis. Therefore, maternal HCMV DNAemia is not a significant risk factor for iatrogenic HCMV transmission to the fetus during amniocentesis.