基于Nanopore平台的测序试剂QzTGS HLA MX9在HLA基因分型中的评价研究

Evaluation of sequencing reagent QzTGS HLA MX9 based on Nanopore platform in HLA genotyping

目的 探讨基于Nanopore平台的测序试剂QzTGS HLA MX9(TGS测序)在人类白细胞抗原(human leukocyte antigen, HLA)基因分型中应用的性能。方法 选取北京市红十字血液中心实验室常规HLA分型检测样本48份,采用PCR-SBT和TGS两种测序法分别对全部样本进行HLA基因分型,比较两种方法的基因分型结果及性能。结果 TGS法和PCR-SBT法对48份样本的HLA-A、-B、-C、-DRB1和-DQB1的基因检测结果,在高分辨水平上完全一致,TGS法可以对全部样本进行直接指定新基因及单一组合分型结果。TGS法在耗时、新基因判定、读长、机器维护、模棱两可结果和实时数据等性能方面优势更明显。结论 与PCR-SBT测序技术相比,基于Nanopore平台的测序试剂QzTGS HLA MX9在指定HLA基因结果方面准确性更高、耗时更短。

Objective To explore the application performance of sequencing reagent QzTGS HLA MX9(TGS sequencing)based on Nanopore platform in human leukocyte antigen(HLA)genotyping.Methods A total of 48 samples of routine HLA typing in Beijing Red Cross Blood Center Laboratory were selected.All samples were genotyped by PCR-SBT and TGS respectively.The genotyping results and performance of the two methods were compared.Results The results of gene detection of HLA-A,-B,-C,-DRB1 and-DQB1 in 48 samples by TGS sequencing method and PCR-SBT sequencing method were completely consistent at high resolution level.TGS sequencing method could directly specify new genes and single combination typing results for all samples.The results of HLA-A,-B,-C,-DRB1 and-DQB1 by TGS and PCR-SBT in 48 samples were completely consistent,and the new genes could be directly designated by TGS and the samples designated as a single combination were 48 cases(100%).TGS had obvious advantages in terms of time-consuming,new gene determination,reading length,machine maintenance,ambiguity,and real-time data.Conclusions Compared with PCR-SBT,QzTGS HLA MX9 based on Nanopore sequencing platform has higher accuracy and less time-consuming.