Complete Next-generation Sequencing: Establishment of Reference Alleles

Overview

Journal Blood Adv

Specialty Hematology

Date 2018 Oct 20

PMID 30337299

Citations 15

Authors

Wajnat A Tounsi

Tracey E Madgett

Neil D Avent

Affiliations

Soon will be listed here.

Abstract

The Rh blood group system (ISBT004) is the second most important blood group after ABO and is the most polymorphic one, with 55 antigens encoded by 2 genes, and This research uses next-generation sequencing (NGS) to sequence the complete gene by amplifying the whole gene using overlapping long-range polymerase chain reaction (LR-PCR) amplicons. The aim was to study different alleles present in the population to establish reference allele sequences by using the analysis of intronic single-nucleotide polymorphisms (SNPs) and their correlation to a specific Rh haplotype. Genomic DNA samples (n = 69) from blood donors of different serologically predicted genotypes including RR (DCe/DCe), RR (DcE/DcE), RR (DCe/DcE), RR (DcE/DCE), Rr (DCe/dce), Rr (DcE/dce), and Rr (Dce/dce) were sequenced and data were then mapped to the human genome reference sequence hg38. We focused on the analysis of hemizygous samples, as these by definition will only have a single copy of For the 69 samples sequenced, different exonic SNPs were detected that correlate with known variants. Multiple intronic SNPs were found in all samples: 21 intronic SNPs were present in all samples indicating their specificity to the haplotype which the hg38 reference sequence encodes. Twenty-three intronic SNPs were found to be R haplotype specific, and 15 were linked to R, R, and R haplotypes. In conclusion, intronic SNPs may represent a novel diagnostic approach to investigate known and novel variants of the and genes, while being a useful approach to establish reference allele sequences.

Citing Articles

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