Oxford Nanopore Technology在HLA-DPB1模棱两可基因分型中的应用初探

doi:10.3969/j.issn.1671-2587.2024.03.011

临床输血与检验 ›› 2024, Vol. 26 ›› Issue (3): 365-370.DOI: 10.3969/j.issn.1671-2587.2024.03.011

Oxford Nanopore Technology在HLA-DPB1模棱两可基因分型中的应用初探

廉雪, 张丹, 温玉洁, 范成艳, 王东梅, 刘娜, 张荣芳, 贾延军

北京市红十字血液中心,北京 100088

收稿日期:2024-03-05 出版日期:2024-06-20 发布日期:2024-06-24
通讯作者: 贾延军,主要从事输血医学方面研究,（E-mail） jiayj321@hotmail.com。
作者简介:廉雪,主要从事HLA分型及血小板抗体研究,（E-mail） 120881191@qq.com。

Application of ONT in Ambiguous Results of the HLA-DPB1 Genotyping

LIAN Xue, ZHANG Dan, WEN Yujie, FAN Chengyan, WANG Dongmei, LIU Na, ZHANG Rongfang, JIA Yanjun

HLA Laboratory, Beijing Red Cross Blood Center, Beijing 100088

Received:2024-03-05 Online:2024-06-20 Published:2024-06-24

摘要/Abstract

摘要： 目的通过对比PCR-SBT、NGS、TGS三种测序试剂在HLA-DPB1基因分型的结果,分析基于ONT平台的TGS测序技术在处理HLA-DPB1基因分型模棱两可情况时的效率、优势及原因。方法采用PCR-SBT、NGS（Illumina平台）、TGS（ONT平台）三种测序试剂分别对48例样本进行HLA-DPB1检测分型。NGS测序和TGS测序以HLA-DPB1命名*后三个区域数字作为高分辨结果进行统计,SBT测序以HLA-DPB1命名*后二个区域数字作为高分辨结果进行统计。结果 48例样本中,TGS没有出现模棱两可结果,全部为唯一结果,而PCR-SBT测序共有45个样本出现模棱两可结果,占比93%（45/48）,NGS共有5个样本出现模棱两可结果,占比10%（5/48）。结论与NGS测序和PCR-SBT测序技术相比,TGS测序可以解决HLA-DPB1基因分型的绝大部分模棱两可情况,提高了测序效率和准确性。

关键词: 人类白细胞抗原, PCR-SBT测序, NGS测序, TGS测序, ONT

Abstract: Objective This study aims to analyze the efficiency, advantages and mechanisms of the ONT platform, a TGS method in ambiguous results of the HLA-DPB1 genotyping in comparison to PCR-SBT and NGS. Methods 48 samples were tested for HLA-DPB1 genotyping by PCR-SBT, NGS（Illumina platform）and TGS (ONT platform). HLA-DPB1 allele was assigned to the third field for NGS and TGS, and the second field for PCR-SBT. Results TGS had no ambiguity with a single allele combination of HLA-DPB1 alleles. Ambiguous results were given in 45 out of 48 samples by PCR-SBT, accounting for 93% (45/48), and 5 out of 48 samples by NGS, accounting for 10% (5/48). Conclusion Compared with NGS and PCR-SBT, TGS can resolve most of the ambiguous combinations and improve the efficiency and accuracy in HLA-DPB1.

Key words: Human leukocyte antigen, PCR-SBT, NGS, TGS, ONT

中图分类号:

R446

廉雪, 张丹, 温玉洁, 范成艳, 王东梅, 刘娜, 张荣芳, 贾延军. Oxford Nanopore Technology在HLA-DPB1模棱两可基因分型中的应用初探[J]. 临床输血与检验, 2024, 26(3): 365-370.

LIAN Xue, ZHANG Dan, WEN Yujie, FAN Chengyan, WANG Dongmei, LIU Na, ZHANG Rongfang, JIA Yanjun. Application of ONT in Ambiguous Results of the HLA-DPB1 Genotyping[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2024, 26(3): 365-370.

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Oxford Nanopore Technology在HLA-DPB1模棱两可基因分型中的应用初探

Application of ONT in Ambiguous Results of the HLA-DPB1 Genotyping

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