Detection of fetal messenger ribonucleic acid in maternal blood to determine fetal RhD status as a strategy for noninvasive prenatal diagnosis☆,☆☆,★,★★
Section snippets
Sample collection
Institutional ethical committee approval was obtained for this study, which complied with national guidelines relating to research in fetal tissues. Three different groups of RhD-negative women were investigated: women in the first trimester at 6 to 12 weeks’ gestation undergoing termination of pregnancy, women in the second trimester (13 to 23 weeks’ gestation) undergoing genetic amniocentesis, and women in the third trimester at ≥36 weeks’ gestation. None received anti-D antenatally. Venous
Results
Reverse transcriptase–PCR generated an amplification product of 567 bp. PCR for DNA generated an amplification product of 286 bp. Thirty-five RhD-negative pregnant women were included in the study; 10 in the first trimester, 10 in the second trimester, and 15 in the third trimester. Table I shows the comparison between reverse transcriptase–PCR and genomic PCR in each of the gestational groups.
Comment
This is the first study to compare reverse transcriptase–PCR with genomic PCR in the same patient group. We found that reverse transcriptase–PCR for detection of fetal RhD messenger RNA in the maternal circulation is more sensitive than PCR for detection of fetal DNA. This is presumably due to a greater template copy number. Although the copy number for RhD is currently unknown, it is likely (as for other expressed sequences) to be many hundred-fold greater than the single strand of RhD DNA
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Cited by (27)
Rhesus and Other Fetomaternal Incompatibilities
2013, Emery and Rimoin's Principles and Practice of Medical GeneticsNoninvasive prenatal detection and selective analysis of cell-free DNA obtained from maternal blood: Evaluation for trisomy 21 and trisomy 18
2012, American Journal of Obstetrics and GynecologyNon-invasive fetal RHD genotyping tests: A systematic review of the quality of reporting of diagnostic accuracy in published studies
2009, European Journal of Obstetrics and Gynecology and Reproductive BiologyCitation Excerpt :Ideally, for RhD NIPD these should include (a) controls of known RhD genotypes and (b) controls for the presence of fetal material. Use of control samples of known RhD type (positive as well as negative) and run at the same time as the index test was reported in only 10 studies [4,5,13,15,25,29,32,37–39]. Only two of these were controls from pregnant women with known RhD-positive or -negative fetuses.
The changing face of haemolytic disease of the newborn
2008, Early Human DevelopmentDiagnostic accuracy of noninvasive fetal Rh genotyping from maternal blood-A meta-analysis
2006, American Journal of Obstetrics and GynecologyCitation Excerpt :There were a total of 3261 samples that were tested for prenatal fetal RhD genotype in peripheral maternal blood. Five studies included more than 1 protocol to determine fetal RhD status from maternal blood.17,21,30,36,44 A total of 183 (183/3261, 5.6%) samples were excluded from the meta-analysis (Figure 1).
Le génotypage RHD prénatal non invasif
2006, Transfusion Clinique et Biologique
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From the Institute of Obstetrics and Gynaecology, Imperial College School of Medicine, Queen Charlotte’s and Chelsea Hospital,a and the Universitats Frauenklinik, Kantonspital.b
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Supported by a North Atlantic Treaty Organization Collaborative Research Grant, an Eden Traveling Fellowship from the Royal College of Obstetricians and Gynaecologists, and a grant from the Swiss National Foundation.
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Reprints not available from the authors.
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