In this study, we assessed the feasibility of fetal RhD genotyping by analysis 
of cell-free fetal DNA(cffDNA) extracted from plasma samples of Rhesus (Rh) 
D-negative pregnant women by using real-time polymerase chain reaction (PCR). 
Fetal genotyping was performed on 30 RhD-negative women between 9 and 39 weeks 
of gestation who were referred to us for invasive testing 
[amniocentesis/chorionic villi sampling (CVS)]. The fetal RHD genotype was 
determined based on real-time PCR method. Exons 7 and 10 of the RHD and SRY 
genes were targeted. Among the pregnant women, 12 were carrying male and 17 were 
carrying female fetuses. Out of 29 pregnant women, 21 had RhD-positive and nine 
had RhD-negative fetuses. One sample (case 12, whose blood group was found to be 
AB Rh [+]) was excluded due to controversial results from repeated serological 
analyses. All prenatal results were in concordance with postnatal RhD status and 
fetal sex without false- positive or -negative results. Performing real-time PCR 
on cffDNA showed accurate, efficient and reliable results, allowing rapid and 
high throughput non invasive determination of fetal sex and RhD status in 
clinical samples.