<scp>MN</scp> typing discrepancies based on <i><scp>GYPA</scp>‐<scp>B</scp>‐<scp>A</scp></i> hybrid

Polin, Helene; Danzer, Martin; Reiter, Anna; Brisner, M.; Gaszner, Waltraud; Weinberger, Johannes; Gabriel, Christian

doi:10.1111/vox.12168

Cited by 6 publications

(6 citation statements)

References 27 publications

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“…Genomic DNA was automatically isolated from ethylenediaminetetraacetate‐anticoagulated blood using MagNa Pure Compact technology (Roche Diagnostics, Mannheim, Germany) according to the manufacturer's instructions. DNA sequencing of RHD , RHCE , GYPA , and GYPB genes was performed as described earlier . Polymerase chain reaction (PCR) amplifications for RHAG exons 1‐10 and adjacent intronic regions were carried out on a LightCyler 480 real‐time instrument (Roche Diagnostics).…”

Section: Methodsmentioning

confidence: 99%

“…DNA sequencing of RHD, RHCE, GYPA, and GYPB genes was performed as described earlier. [9][10][11] Polymerase chain reaction (PCR) amplifications for RHAG exons 1-10 and adjacent intronic regions were carried out on a LightCyler 480 real-time instrument (Roche Diagnostics). The program consisted of an initial denaturation of 10 min at 958C, followed by 35 amplification cycles at 958C for 1 s, 618C for 5 s and 728C for 30 s. Each PCR reaction contained 2.5 mM MgCl2, 0.25 mM of each primer published before, 15 13 LightCycler SYBR Green I Master Mix (Roche Diagnostics), and 10 ng of DNA in a final volume of 10 mL.…”

Section: Genomic Dna Studymentioning

confidence: 99%

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Compound heterozygosity of two novel RHAG alleles leads to a considerable disruption of the Rh complex

et al. 2016

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Section: Methodsmentioning

confidence: 99%

Section: Genomic Dna Studymentioning

confidence: 99%

Compound heterozygosity of two novel RHAG alleles leads to a considerable disruption of the Rh complex

et al. 2016

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“…The PCR reaction was performed under the following conditions: activation at 95°C for 2 min, 30 cycles of denaturation at 95°C for 30 s, annealing at 56°C for 30 s, and extension at 72°C for 30 s, followed by a final extension at 72°C for 5 min. The coding regions of GYPA were amplified using the primers described before, 32 and the PCR procedure is similar with GYPB cDNA amplification except that the annealing temperature was 60°C. PCR products were separated on a 3% agarose gel by electrophoresis for 3 h at 100 V. The electrophoresis bands were cut from the gel and purified using a Gel Extraction Kit (OMEGA) in accordance with manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Molecular genetic analysis of Mi^a‐positive hybrid glycophorins revealed two novel alleles of GP.Vw and multiple variant transcripts of GYPB existing in both the homozygous GP.Mur and wild‐type GPB individuals

et al. 2021

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Background The hybrid glycophorins of MNS blood group system express a series of low incidence antigens including Mia, which are commonly found in Southeast Asian populations. In this study, the molecular basis of Mia‐positive hybrid glycophorins was firstly clarified in the Chinese Southern Han population. RNA transcripts of GYPB gene in the homozygous GP.Mur individuals were also analyzed. Study design and methods DNAs were extracted from the whole blood samples of 111 Mia‐positive donors. Then, high‐resolution melting (HRM) analysis for GYP(B‐A‐B) was used to analyze the genotypes. Sequencing of GYPB pseudoexon 3 was conducted in the samples with variant melting curves. TA‐cloning and subsequent sequencing of GYPA exons 2–4 were performed in the Mia‐positive samples with normal GYPB/GYPB genotype by HRM. The transcript analysis of GYPB was conducted in homozygous GP.Mur and wild‐type glycophorin B (GPB) individuals using RNA extracted from the cultured erythroblast. Results The heterozygous GYP*Mur/GYPB (n = 101), homozygous GYP*Mur/GYP*Mur (n = 7) including one novel GYP*Mur allele with an extra GYPA/GYPE specific nucleotide substitution (c.229+110A>T), heterozygous GYP*Bun/GYPB (n = 1) and GYP*Vw/GYPA (n = 2) with two novel GYP*Vw alleles were identified. RNA transcript analysis revealed multiple transcripts of GYPB existing in both homozygous GP.Mur and normal GPB individuals. Conclusion The results showed the genetic diversity of hybrid glycophorins in the Chinese population. Besides, the successful analysis of GYPB transcripts indicates that the cultured erythroblast is a good source for RNA transcript analysis for the protein only expressed on the red blood cells.

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Section: Introductionmentioning

confidence: 99%

“…The GYPA, GYPB, and GYPE genes are evolutionarily related, with at least 95% sequence homology between them resulting from duplication events, whereby GYPB evolved from GYPA, and GYPE evolved from GYPB. 9,25,26 Recently, a large structural variant in the chromosome 4 GYP region that gives rise to the Dantu glycophorin (DUP4) has been associated with about a 40% reduction in risk for severe malaria. 9,27 Interestingly, this Dantu variant was found predominantly in East African populations, but there are many other common structural variants across all the West and East African populations that have been studied.…”

Section: Introductionmentioning

confidence: 99%

High-throughput genotyping assays for identification of glycophorin B deletion variants in population studies

Amuzu

Rockett

Leffler

et al. 2020

Exp Biol Med (Maywood)

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Glycophorins are the most abundant sialoglycoproteins on the surface of human erythrocyte membranes. Genetic variation in glycophorin region of human chromosome 4 (containing GYPA, GYPB, and GYPE genes) is of interest because the gene products serve as receptors for pathogens of major public health interest, including Plasmodium sp., Babesia sp., Influenza virus, Vibrio cholerae El Tor Hemolysin, and Escherichia coli. A large structural rearrangement and hybrid glycophorin variant, known as Dantu, which was identified in East African populations, has been linked with a 40% reduction in risk for severe malaria. Apart from Dantu, other large structural variants exist, with the most common being deletion of the whole GYPB gene and its surrounding region, resulting in multiple different deletion forms. In West Africa particularly, these deletions are estimated to account for between 5 and 15% of the variation in different populations, mostly attributed to the forms known as DEL1 and DEL2. Due to the lack of specific variant assays, little is known of the distribution of these variants. Here, we report a modification of a previous GYPB DEL1 assay and the development of a novel GYPB DEL2 assay as high-throughput PCR-RFLP assays, as well as the identification of the crossover/breakpoint for GYPB DEL2. Using 393 samples from three study sites in Ghana as well as samples from HapMap and 1000 G projects for validation, we show that our assays are sensitive and reliable for genotyping GYPB DEL1 and DEL2. To the best of our knowledge, this is the first report of such high-throughput genotyping assays by PCR-RFLP for identifying specific GYPB deletion types in populations. These assays will enable better identification of GYPB deletions for large genetic association studies and functional experiments to understand the role of this gene cluster region in susceptibility to malaria and other diseases.

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MN typing discrepancies based on GYPA‐B‐A hybrid

Abstract: The results document a GYPA-B-A hybrid gene, probably produced via a single unequal homologous recombination event. A segmental transfer of GYPB seems most likely accounting for the allelic dropout.

Cited by 6 publications

References 27 publications

Compound heterozygosity of two novel RHAG alleles leads to a considerable disruption of the Rh complex

Compound heterozygosity of two novel RHAG alleles leads to a considerable disruption of the Rh complex

Molecular genetic analysis of Mi^a‐positive hybrid glycophorins revealed two novel alleles of GP.Vw and multiple variant transcripts of GYPB existing in both the homozygous GP.Mur and wild‐type GPB individuals

High-throughput genotyping assays for identification of glycophorin B deletion variants in population studies

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MN typing discrepancies based on GYPA‐B‐A hybrid

Abstract: The results document a GYPA-B-A hybrid gene, probably produced via a single unequal homologous recombination event. A segmental transfer of GYPB seems most likely accounting for the allelic dropout.

Cited by 6 publications

References 27 publications

Compound heterozygosity of two novel RHAG alleles leads to a considerable disruption of the Rh complex

Compound heterozygosity of two novel RHAG alleles leads to a considerable disruption of the Rh complex

Molecular genetic analysis of Mia‐positive hybrid glycophorins revealed two novel alleles of GP.Vw and multiple variant transcripts of GYPB existing in both the homozygous GP.Mur and wild‐type GPB individuals

High-throughput genotyping assays for identification of glycophorin B deletion variants in population studies

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Molecular genetic analysis of Mi^a‐positive hybrid glycophorins revealed two novel alleles of GP.Vw and multiple variant transcripts of GYPB existing in both the homozygous GP.Mur and wild‐type GPB individuals