【摘要】 目的 AllType^TM Fastplex^TM NGS试剂增加了HLA-Ⅱ类基因1号外显子序列分析，本文旨在评价该试剂基于Ion Torrent S5平台进行HLA基因分型的性能情况。方法 利用AllType^TM Fastplex^TM NGS试剂检测HLA-A、-B、-C、-DRB1、-DQB1、-DPB1位点，使用TSV 2.0.0软件指定NGS法的HLA基因型。采用聚合酶链反应-序列分析法（PCR-SBT）复核部分标本进行质量控制。结果 94份标本NGS法检测HLA-A、-B、-C、-DRB1、-DQB1、-DPB1位点，指定为单一组合分型结果为58份，占61.70%。结果分析中软件提示至少有一个等位基因报警的组合有432个，占76.60%。共有5个等位基因组合，报警存在外显子区域碱基错配，经PCR-SBT方法复核发现均为NGS软件错误的判定。随机抽取16份标本，NGS法和PCR-SBT方法检测HLA-A、-B、-C、-DRB1、-DQB1、-DPB1基因结果在高分辨水平上完全一致。增加HLA-Ⅱ类基因1号外显子序列，有效解决了DRB1*12:01:01/12:10、DRB1*15:02:01/15:140/15:149、DPB1*13:01/107:01组合的指定。结论 AllType^TM Fastplex^TM NGS试剂增加分析HLA-Ⅱ类基因1号外显子，提升HLA分型指定能力。HLA基因型结果指定中应注意软件分析报警信息的处理，必要时采用其他方法进行验证，以保证分型结果准确性。更多还原

【Abstract】 Objective AllType^TM Fastplex^TM NGS kit can analyze the exon 1 sequence of the HLA class II gene. This study aims to evaluate the performance of this reagent for HLA genotyping based on Ion Torrent S5 platform. Methods The HLA-A,-B,-C,-DRB1,-DQB1,-DPB1 loci were genotyped using the AllType^TM Fastplex^TM NGS kit, and the HLA genotype was assigned by TSV 2.0.0 software. Ploymerase chain reaction sequence-based typing（PCR-SBT） was used to recheck some samples for quality control. Results The HLA-A,-B,-C,-DRB1,-DQB1,-DPB1 loci were detected in 94 specimens by the NGS method. Among them, a unique alleles combination was assigned in the 58 specimens, accounting for 61.70%. 432 alleles combinations with at least one allele alarm were found in the result analysis, accounting for 76.60%. There are 5 alleles combinations with bases mismatch in exon regions alarm, all of them were error in the software analysis by PCR-SBT retest. The results of HLA-A,-B,-C,-DRB1,-DQB1,-DPB1 loci in the 16 randomly specimens were completely consistent at high resolution level between the NGS method and PCR-SBT method. The HLA-DRB1*12:01:01/12:10,-DRB1*15:02:01/15:140/15:149 and-DPB1*13:01/107:01 combinations were effectively discriminated by addition of exon 1 sequence of HLA class Ⅱ gene. Conclusion AllType^TM Fastplex^TM NGS kit can analyze the exon 1 sequence of the HLA class Ⅱ gene and improves the ability of HLA genotype assignment. It should be paid attention to process software analysis alarm information in the HLA genotype assignment procedure, and other methods should be used for verification if necessary to ensure the accuracy of genotyping results.更多还原