小年 (2024-07-31 15:47):
#paper DOI: 10.1016/j.csbj.2024.03.030 A novel framework for human leukocyte antigen (HLA) genotyping using probe capture - based targeted next - generation sequencing and computational analysis 这篇文章介绍了一种利用基于探针捕获的靶向下一代测序和计算分析进行人类白细胞抗原(HLA)基因分型的分析流程,研究团队没有使用常用的IPD-IMGT/HLA 数据库做参考而是使用了人类泛基因组参考联盟(HPRC)资源作为HLA参考的基准,丰富了HLA参考数据库,为解决传统参考存在的问题提供了新的思路。在算法方面该团队使用了五个开源软件工具(OptiType、HLA - VBseq、HISAT - genotype、SpecHLA和T1K)作比较,虽然没有单一一种软件可以做到分型100%正确,但结合使用T1K、DRAGEN和QzType三个工具进行联合分析能使HLA基因的准确率达到100%。该研究证明了HLA分型基于探针捕获的靶向测序的有效性,特别是结合集成软件分析方法,能够提高HLA分型的准确性。
A novel framework for human leukocyte antigen (HLA) genotyping using probe capture-based targeted next-generation sequencing and computational analysis
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Abstract:
Human leukocyte antigen (HLA) genes play pivotal roles in numerous immunological applications. Given the immense number of polymorphisms, achieving accurate high-throughput HLA typing remains challenging. This study aimed to harness the human pan-genome reference consortium (HPRC) resources as a potential benchmark for HLA reference materials. We meticulously annotated specific four field-resolution alleles for 11 HLA genes (HLA, , , , , , , , , and ) from 44 high-quality HPRC personal genome assemblies. For sequencing, we crafted HLA-specific probes and conducted capture-based targeted sequencing of the genomic DNA of the HPRC cohort, ensuring focused and comprehensive coverage of the HLA region of interest. We used publicly available short-read whole-genome sequencing (WGS) data from identical samples to offer a comparative perspective. To decipher the vast amount of sequencing data, we employed seven distinct software tools: OptiType, HLA-VBseq, HISAT genotype, SpecHLA, T1K, QzType, and DRAGEN. Each tool offers unique capabilities and algorithms for HLA genotyping, allowing comprehensive analysis and validation of the results. We then compared these results with benchmarks derived from personal genome assemblies. Our findings present a comprehensive four-field-resolution HLA allele annotation for 44 HPRC samples. Significantly, our innovative targeted next-generation sequencing (NGS) approach for HLA genes showed superior accuracy compared with conventional short-read WGS. An integrated analysis involving QzType, T1K, and DRAGEN was developed, achieving 100% accuracy for all 11 HLA genes. In conclusion, our study highlighted the combination of targeted short-read sequencing and astute computational analysis as a robust approach for HLA genotyping. Furthermore, the HPRC cohort has emerged as a valuable assembly-based reference in this realm.
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