基于Illumina Miseq测序技术的8000份标本HLA分型检测结果分析

发布时间：2018-02-09 05:41

本文关键词： HLA分型 NGS测序技术 PCR-SBT 高分辨率中华骨髓库　出处：《中国输血杂志》2016年09期 　论文类型：期刊论文

【摘要】：目的利用Illumina Miseq二代测序(NGS)技术进行HLA分型工作并分析其结果的准确性。方法利用Illumina Miseq二代测序技术,对8 000人份血液标本进行HLA-A,HLA-B和HLA-DRB1位点的分型工作。为验证二代测序技术的准确性,同时选取1 000人份血液标本进行一代测序(PCR-SBT)分型实验,比较两种测序方法所得分型结果。结果分型结果显示PCR-NGS HLA分型技术获得的数据与一代测序PCR-SBT技术数据完全吻合,且利用NGS测序技术的8 000人份标本中高分辨率结果 7 908份(含G族1 058人份),中分辨率结果 92份,高分辨率比例达到98.85%。结论利用NGS测序技术进行HLA分型可以大大减少实验工作量,提高高分辨率结果比例,更容易对罕见型和新等位基因进行鉴定,在工作量大,时间紧迫时相对于传统的一代测序分型有明显优势。
[Abstract]:Objective to use Illumina Miseq second-generation sequencing technique to carry out HLA typing and analyze the accuracy of the results. Methods Illumina Miseq second-generation sequencing technique was used. In order to verify the accuracy of the second generation sequencing technique, 1 000 blood samples were selected to carry out a generation sequencing PCR-SBT typing experiment, which was used to type the HLA-AG HLA-B and HLA-DRB1 loci in 8 000 blood samples, in order to verify the accuracy of the second generation sequencing technique, 1 000 blood samples were selected for a generation sequencing PCR-SBT typing experiment. The results showed that the data obtained by the PCR-NGS HLA typing method were in good agreement with the data obtained by the first generation sequencing PCR-SBT technique. Among the 8 000 samples by NGS sequencing technique, 7 908 samples were obtained with high resolution (including 1 058 samples of G group, 92 with medium resolution results). Conclusion using NGS sequencing technique for HLA typing can greatly reduce the workload of experiments, increase the proportion of high resolution results, and make it easier to identify rare and new alleles. When time is short, it has obvious advantage over traditional generation sequencing typing.
【作者单位】：河北省血液中心;北京诺诗康瀛医学科技有限公司;
【分类号】：R440

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