一例HLA-A新等位基因A*2459的测序分析

本研究探讨HLA新等位基因HLA-A*2459的分子机制。样本DNA抽提采用PEL-FREEZ抽提试剂盒,利用单链特异性引物PCR方法扩增先证者HLA-A基因的第2-4外显子,PCR产物直接进行第2、3、4外显子双向测序分析。结果显示:先证者样本中存在2个HLA-A等位基因,其中1个等位基因为HLA-A*1101,另1个经Blast验证其为新的等位基因,新的等位基因序列已递交GenBank(DQ313255,DQ313256,DQ313257)。与最接近的HLA-A*24020101等位基因序列相比,新的等位基因仅在第3外显子上有1个核苷酸不同,即第527位T→C;这导致氨基酸第152位Val→Ala。结论:该等位基因为新的HLA-A等位基因,被世界卫生组织HLA因子命名委员会正式命名为HLA-A*2459。

Sequence Analysis of A Novel HLA-A*2459 Allele

This study was aimed to investigate the molecular genetics basis of a novel allele HLA-A*2459 in Chinese population. DNA was extracted from whole blood by PEL-FREEZ DNA extraction kit. The amplification of HLA-A exons 2-4 of the proband was preformed by allele specific primer PCR and the amplified product was sequenced bidirectionally with primers. The sequencing results showed HLA-A alleles of the proband as A*1101 and the novel allele. The sequences of the novel allele have been submitted to GenBank(DQ313255,DQ313256,DQ313257). After Blast HLA analysis, the novel allele showed only one nucleotide differences with HLA-A*24020101 at nucleotide position 527 T to C in exon 3. This results in an amino acid changes from Val to Ala at codon 152. In conclusion, this allele is a novel one and has been officially named HLA-A*2459 by the WHO Nomenclature Committee.