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Hemolytic  disease of the newborn is a clinical condition in which maternal and paternal  Rh blood group antigens are incompatible and the mother is negative for the  antigen whereas the father is positive. Analysis of fetal cells recovered from  maternal plasma can provide a highly sensitive prenatal diagnosis. The fetal RHD gene in plasma DNA is detected by real-time PCR amplification of two  different segments of the RHD gene (exons 7 and 10). Each amplicon is  revealed with specific probes. We examined 40 female blood samples to verify  the specificity of RHD exons (7 and 10) amplified by real-time PCR. Thirty  fetuses were predicted to be RHD-positive based on analysis of plasma DNA.  Seven fetuses were predicted to be RHD-negative. One fetus was negative for RHD  on exon 10, and positive for RHD on exon 7 (early gestation age); two fetuses  were RHD-negative on exon 7, and RHD-positive on exon 10 (RHD-CE-D(s) or RHDΨ), indicative of a  maternal RHD allele. We conclude that it is necessary to analyze at least two  exon regions in the RHD gene.

Detection of fetal RHD pseudogene (RHDΨ) and hybrid RHD-CE-Ds from RHD-negative pregnant women with a free DNA fetal kit