Background: The most common large-deletion RHD allele (RHD*01N.01) includes the entire coding sequence, intervening regions and untranslated regions. The rest of large-deletion RHD alleles reported to-date consist of single-exon deletions, such as RHD*01N.67 which includes exon 1. Materials and Methods: Samples from two donors with RhD-negative serology yielded unclear or inconclusive results when subject to confirmatory testing on RHD genotyping arrays. To determine their RHD genotypes, genomic DNA was analyzed with a combination of allele-specific PCR, long-range PCR, Sanger sequencing, and next-generation sequencing assays. Results: Allele-specific PCR failed to detect products for RHD exons 1 to 3 in one sample and RHD exons 1 to 5 in the other. A quantitative next-generation sequencing assay confirmed deletion of exons 1 to 3 and 1 to 5 respectively, and detected the absence of an RHD gene in trans in both samples. Long-range PCR and Sanger sequencing enabled identification of the breakpoints for both alleles. Both deletions start within the 5′ Rhesus box (upstream of the identity region for the 1-to-3 deletion, downstream of it for the 1-to-5 deletion), and end within introns. Conclusions: Resolution of unclear or inconclusive results from targeted genotyping arrays often leads to the discovery of new alleles. The 5′ Rhesus box may be a hot spot for genetic recombination events, such as the large deletions described in this report. © 2020 AABB

Two new RHD alleles with deletions spanning multiple exons / Matteocci, A; Monge-Ruiz, J; Stef, M; Apraiz, ; I., Herrera-del-Val L; Mancuso, T; Fennell, K; Lopez, M; Larizgoitia-Martin, Y; Nespoli, G; Rubia-Tejero, M; Collaretti, A; Pierelli, L; Ochoa-Garay, G.. - In: TRANSFUSION. - ISSN 0041-1132. - 61:3(2021), pp. 682-686. [10.1111/trf.16199]

Two new RHD alleles with deletions spanning multiple exons

Pierelli L;
2021

Abstract

Background: The most common large-deletion RHD allele (RHD*01N.01) includes the entire coding sequence, intervening regions and untranslated regions. The rest of large-deletion RHD alleles reported to-date consist of single-exon deletions, such as RHD*01N.67 which includes exon 1. Materials and Methods: Samples from two donors with RhD-negative serology yielded unclear or inconclusive results when subject to confirmatory testing on RHD genotyping arrays. To determine their RHD genotypes, genomic DNA was analyzed with a combination of allele-specific PCR, long-range PCR, Sanger sequencing, and next-generation sequencing assays. Results: Allele-specific PCR failed to detect products for RHD exons 1 to 3 in one sample and RHD exons 1 to 5 in the other. A quantitative next-generation sequencing assay confirmed deletion of exons 1 to 3 and 1 to 5 respectively, and detected the absence of an RHD gene in trans in both samples. Long-range PCR and Sanger sequencing enabled identification of the breakpoints for both alleles. Both deletions start within the 5′ Rhesus box (upstream of the identity region for the 1-to-3 deletion, downstream of it for the 1-to-5 deletion), and end within introns. Conclusions: Resolution of unclear or inconclusive results from targeted genotyping arrays often leads to the discovery of new alleles. The 5′ Rhesus box may be a hot spot for genetic recombination events, such as the large deletions described in this report. © 2020 AABB
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1577175
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