Faculty of Medicine and Dentistry, School of Biomedical Sciences, University of Plymouth, Plymouth, United Kingdom; and.
School of Medical Technology, King Abdulaziz University, Jeddah, Saudi Arabia.
Blood Adv. 2018 Oct 23;2(20):2713-2723. doi: 10.1182/bloodadvances.2018017871.
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.
Rh 血型系统（ISBT004）是仅次于 ABO 血型系统的第二大重要血型系统，也是最具多态性的血型系统，其 55 种抗原由 2 个基因编码。本研究采用下一代测序（NGS）技术，通过重叠长程聚合酶链反应（LR-PCR）扩增整个基因，对完整的 基因进行测序。目的是研究人群中存在的不同 等位基因，通过分析内含子单核苷酸多态性（SNP）及其与特定 Rh 单倍型的相关性，建立参考 等位基因序列。从不同血清学预测基因型的献血者（n = 69）基因组 DNA 样本中（RR（DCe/DCe）、RR（DcE/DcE）、RR（DCe/DcE）、RR（DcE/DCE）、Rr（DCe/dce）、Rr（DcE/dce）和 Rr（Dce/dce））中扩增出整个基因，然后将数据映射到人类基因组参考序列 hg38 上。我们重点分析了半合子样本，因为这些样本根据定义只会有一个 的单拷贝。在对 69 个测序样本的分析中，检测到与已知变体相关的不同外显子 SNP。在所有样本中均发现多个内含子 SNP：21 个内含子 SNP 存在于所有样本中，表明其与 hg38 参考序列编码的 单倍型具有特异性。23 个内含子 SNP 是 R 单倍型特异性的，15 个 SNP 与 R、R 和 R 单倍型相关。总之，内含子 SNP 可能代表一种新的诊断方法，可用于研究 和 基因的已知和新型变体，同时也是建立参考 等位基因序列的有用方法。
