An Additional Pre-amplification Step for the Early Determination of Fetal RHD from Maternal Plasma

Dovč-Drnovšek, Tadeja; Toplak, Nataša; Bricl, Irena; Blejec, Tanja; Kovač, Minka; Rožman, Primož

doi:10.1007/978-90-481-9382-0_20

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“…Cell-free DNA (cf-DNA) was extracted from 400 μl of plasma using the Biorobot EZ1 workstation and the commercial kit EZ1 Virus Mini Kit v2.0 according to the manufacturer's protocol (Qiagen) [33]. Cf-DNA was eluted in 60 μl of elution buffer.…”

Section: Selection Of Appropriate Rhd Regions For Pcrmentioning

confidence: 99%

Reliable Determination of Fetal RhD Status by RHD Genotyping from Maternal Plasma

Dovč-Drnovšek

Klemenc

Toplak

et al. 2013

Transfus Med Hemother

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Background: Immunoprophylaxis with IgG anti-D is a standard prevention of hemolytic disease of the fetus and newborn. Fetal Rhesus D (RhD) blood group genotyping from maternal plasma of RhD-negative pregnant women allows targeted prophylaxis with IgG anti-D in RhD-positive pregnancies only. We set up a reliable protocol for prenatal RHD genotyping. Methods: 153 pregnant Caucasian RhD-negative women were tested in the 27th week (range 7–38th week) of pregnancy. 18 of them were alloimmunized to the RhD antigen. The fetal RHD genotype was determined based on an automated DNA extraction and real-time polymerase chain reaction method. Intron 4 and exons 5, 7 and 10 of the RHD gene and the SRY gene were targeted. Results: The fetal RhD status and gender was 100% correctly predicted in all 153 pregnancies (55 RhD-positive males, 45 RhD-positive females; 23 RhD-negative males, 30 RhD-negative females). Conclusion: The accuracy and applicability of our protocol for non-invasive fetal RhD determination allows the correct management of RhD-incompatible pregnancies. Our protocol could prevent unnecessary immunoprophylaxis in 53 of 153 cases. We therefore recommend that non-invasive fetal RHD genotyping is introduced as an obligatory part of prenatal screening.

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Section: Selection Of Appropriate Rhd Regions For Pcrmentioning

confidence: 99%