Characterization of novel RHD alleles: relationship between phenotype, genotype, and trimeric architecture

Silvy, Monique; Chapel‐Fernandes, Sylvie; Callebaut, Isabelle; Beley, Sophie; Durousseau, Cécile; Simon, Sophie; Lauroua, P.; Dubosc-Marchenay, Nadine; Babault, Catherine; Mouchet, C.; Ferrera, Virginie; Chiaroni, Jacques; Bailly, Pascal

doi:10.1111/j.1537-2995.2011.03544.x

Cited by 34 publications

(46 citation statements)

References 49 publications

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“…Model of the 3D structure of human RhD and heterotrimer RhAG‐RhD‐RhD was previously published . Visualization of the potential effect of variation on the stability of the heterotrimer was carried out with a freely available program (UCSF Chimera) .…”

Section: Methodsmentioning

confidence: 99%

Functional analysis of novel RHD variants: splicing disruption is likely to be a common mechanism of variant D phenotype

Raud

Gourlaouen

et al. 2019

Transfusion

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BACKGROUND We previously showed that several variations in the RHD gene, including synonymous changes, can be classified as splice site variants and may play a direct role in D variant phenotype expression. We sought to extend our study to additional candidates, notably in the first and last exons of the gene, by engineering a novel universal splice reporting vector, i.e., minigene. STUDY DESIGN AND METHODS Our previous plasmid construct was modified to allow subcloning of any exon(s) of interest for assessing effect of variations on splicing. Seventeen novel and/or uncharacterized variations of the RHD gene were selected for the study and tested in our novel model. RESULTS We engineered and validated a novel universal minigene for assessing virtually any variations of interest for splicing defect. Of the 17 variants tested in the novel model, 11 were shown to alter splicing either totally or partially, including the silent c.1065C>T variation, which induces major skipping of exon 7, and may therefore be responsible for reducing D antigen expression. We also showed that while all three missense variations c.1154G>C, c.1154G>T, and c.1154G>A in exon 9 are splice site variants, splicing is differentially altered and D‐negative phenotype observed in the presence of the latter substitution is likely due to a defect in RhD protein folding. CONCLUSION Overall, we hypothesize that splicing alteration is likely to be a common mechanism of D phenotype variation that has been underestimated so far. Further large‐scale studies are necessary to demonstrate this statement definitely.

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Section: Methodsmentioning

confidence: 99%

Functional analysis of novel RHD variants: splicing disruption is likely to be a common mechanism of variant D phenotype

Raud

Gourlaouen

et al. 2019

Transfusion

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“…This approach was also recommended by other authors [2,24], who suggested including exons 4 or 5 and exons 7 or 10 so as to avoid wrong fetal RHD genotyping due to rare Rh blood group polymorphisms [25]. These exons depend on different mutations or DNA sequence exchanges between RHD and RHCE , the two homologous genes that are responsible for the RhD phenotype.…”

Section: Discussionmentioning

confidence: 99%

Non-Invasive Prenatal RHD Genotyping Using Cell-Free Fetal DNA from Maternal Plasma: An Italian Experience

Picchiassi

Renzo

Tarquini

et al. 2014

Transfus Med Hemother

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Background: This study assessed the diagnostic accuracy of a non-invasive approach to fetal RHD genotyping using cell-free fetal DNA in maternal plasma and a combination of methodological strategies. Methods: Real-time PCR (qPCR) was performed on 216 RhD-negative women between weeks 10+0 and 14+6 of gestation (1st qPCR). qPCR was repeated (2nd qPCR) to increase the amount of each sample for analysis, on 95 plasma aliquots that were available from first trimester blood collection (group 1) and on 13 samples that were collected between weeks 18+0 and 25+6 of gestation (group 2). qPCR was specific for exons 5 and 7 of the RHD gene (RHD5 and RHD7). The results were interpreted according to the number of positive replicates of both exons. Results: 1st qPCR: diagnostic accuracy was of 93.3%. Diagnostic accuracy increased from 90.5% (1st qPCR) to 93.7% (2nd qPCR) in group 1 and from 84.6% (1st qPCR) to 92.3% (2nd qPCR) in group 2. These increments were not statistically significant. Conclusion: Our approach to RHD genotyping in early pregnancy yielded high diagnostic accuracy. Increasing the amount of DNA analyzed in each sample did not improve significantly the diagnostic accuracy of the test.

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“…Both DAR and weak D -4.2 phenotypes are associated with anti -D production, emphasising the diffi culties of the partial D/weak D terminology. Another variation is DAR with an additional Thr203Ala [77] . Another variation is DAR with an additional Thr203Ala [77] .…”

Section: Dar Dare and Weak D Type 42mentioning

confidence: 99%

“…Any further attempts to analyse the nature of the D epitopes must take into account the architecture of the RhD -RhAG heterotrimers [77] .…”

Section: Variants and Transfusion Practicementioning

confidence: 99%

Rh and RHAG Blood Group Systems

Daniels

2013

Human Blood Groups

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Characterization of novel RHD alleles: relationship between phenotype, genotype, and trimeric architecture

Abstract: Although the frequency of the new RHD alleles presented herein is low, their phenotypic and genotypic description adds to the repertoire of reported RHD alleles. These data can be useful for optimization of molecular screening tools.

Cited by 34 publications

References 49 publications

Functional analysis of novel RHD variants: splicing disruption is likely to be a common mechanism of variant D phenotype

Functional analysis of novel RHD variants: splicing disruption is likely to be a common mechanism of variant D phenotype

Non-Invasive Prenatal <b><i>RHD</i></b> Genotyping Using Cell-Free Fetal DNA from Maternal Plasma: An Italian Experience

Rh and RHAG Blood Group Systems

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