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利用纳米孔测序技术,在一名来自中国骨髓捐献者计划的志愿者中发现了一种新型 HLA-A*11:335 等位基因,该基因是由 HLA-A*11:01:01:01/126 和 HLA-H*02:07/14/18 等位基因之间罕见的基因间重组导致的。

Characterization of a novel HLA-A*11:335 allele resulting from a rare interlocus recombination involving HLA-A*11:01:01:01/126 and HLA-H*02:07/14/18 alleles with nanopore sequencing, in a volunteer from the China Marrow Donor Program.

机构信息

The Key Laboratory of Geriatrics, Beijing Institute of Geriatrics, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing Hospital/National Center of Gerontology of National Health Commission, No. 1 DaHua Road, Dong Dan, Beijing, People's Republic of China.

GenDx, Genome Diagnostics B.V., Utrecht, the Netherlands.

出版信息

BMC Med Genomics. 2022 Mar 16;15(1):58. doi: 10.1186/s12920-022-01176-1.

Abstract

BACKGROUND

The major histocompatibility complex (MHC) in humans includes three classical class I loci (A, B, and C), which are important biomarkers for the transplantation of organs and hematopoietic stem cells. In the MHC, polymorphism is known to be extremely high while interlocus recombination is rare. We report a rare interlocus recombination between HLA-A and HLA-H, which was analyzed using next generation sequencing and nanopore sequencing.

METHODS

In the sample, the genotypes of HLA-A, B, C, DRB1, and DQB1 were firstly determined using the methods of sequence-specific primer, sequence-specific oligonucleotide, Sanger's sequencing, and NGS; however, HLA-A could not be phased. Nanopore sequencing was finally utilized to distinguish the sequence of the novel allele.

RESULTS

Finally, the novel HLA-A11:335 allele was identified as an interlocus recombination involving HLA-A11:01:01:01/126 and HLA-H*02:07/14/18 alleles; this was mainly achieved by nanopore sequencing.

CONCLUSIONS

The identification of the interlocus recombination indicated that nanopore sequencing can be helpful in the characterization of novel alleles with complex rearrangements. Interlocus recombination has been identified as one of the mechanisms involved in the generation of novel HLA alleles.

摘要

背景

人类主要组织相容性复合体(MHC)包括三个经典的 I 类基因座(A、B 和 C),这些基因座是器官和造血干细胞移植的重要生物标志物。在 MHC 中,多态性极高,而基因座间重组罕见。我们报告了 HLA-A 和 HLA-H 之间的罕见基因座间重组,该重组通过下一代测序和纳米孔测序进行分析。

方法

在该样本中,首先采用序列特异性引物、序列特异性寡核苷酸、Sanger 测序和 NGS 方法确定 HLA-A、B、C、DRB1 和 DQB1 的基因型;然而,HLA-A 无法定相。最后利用纳米孔测序来区分新等位基因的序列。

结果

最终确定新型 HLA-A11:335 等位基因为 HLA-A11:01:01:01/126 和 HLA-H*02:07/14/18 等位基因间的重组,这主要是通过纳米孔测序实现的。

结论

该基因座间重组的鉴定表明,纳米孔测序有助于对具有复杂重排的新型等位基因进行特征描述。基因座间重组已被确定为产生新 HLA 等位基因的机制之一。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da4d/8925214/7a3e3377419c/12920_2022_1176_Fig1_HTML.jpg

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