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

作　　者：廉雪 张丹 温玉洁[1] 范成艳 王东梅[1] 刘娜[1] 张荣芳 贾延军[1] 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)

机构地区：[1]北京市红十字血液中心,北京100088

出　　处：《临床输血与检验》2024年第3期365-370,共6页Journal of Clinical Transfusion and Laboratory Medicine

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。Objective This study aims to analyze the efficiency,advantages and mechanisms of the ONT platform,a TGS method in ambiguous results of the HLA-DPB 1 genotyping in comparison to PCR-SBT and NGS.Methods 48 samples were tested for HLA-DPB 1 genotyping by PCR-SBT,NGS(Illumina platform)and TGS(ONT platform).HLA-DPB 1 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-DPB 1 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-DPB 1.

关 键 词：人类白细胞抗原 PCR-SBT测序 NGS测序 TGS测序 ONT 

分 类 号：R446[医药卫生—诊断学]