CAJ | 학술논문

目的鉴定中国人群人类白细胞抗原(human leukocyte antigen,HLA)A*02:251新等位基因,分析新等位基因遗传特征.方法采用聚合酶链反应-测序分型法(polymerase chain reaction-sequence based typing,PCR-SBT)对组织配型健康供、患者进行HLA基因分型,发现先证者核苷酸杂合序列与已知序列不匹配,不能指定先证者HLA等位基因型,对先证者DNA扩增HLA-A位点第2～4外显子,PCR产物经克隆到PMD18-T质粒载体中以获得单链核苷酸序列,对克隆所得产物进行HLA-A基因的第2～4外显子双向测序分析.结果发现先证者的一个HLA-A*02:06:01基因被确认,而另一个HLA-A基因为新等位基因,其序列被GenBank接受(编号为HM245348).新等位基因序列通过IMGT/HLA 数据库BLAST,与最相近的A*02:01:01:01相比,在第3外显子上有1个核苷酸的不同,即第383位 G>C,密码子 128 GAG→GAC,氨基酸由谷氨酸(Glu)→天门冬氨酸(Asp).供、患者HLA-A、B、C、DQB1位点等位基因不匹配.结论该等位基因为新的HLA-A*02:251等位基因.中国人群HLA-A 位点第3外显子核苷酸序列存在多态性.
목적 감정중국인군인류백세포항원(human leukocyte antigen,HLA)A*02:251신등위기인,분석신등위기인유전특정.방법 채용취합매련반응-측서분형법(polymerase chain reaction-sequence based typing,PCR-SBT)대조직배형건강공、환자진행HLA기인분형,발현선증자핵감산잡합서렬여이지서렬불필배,불능지정선증자HLA등위기인형,대선증자DNA확증HLA-A위점제2～4외현자,PCR산물경극륭도PMD18-T질립재체중이획득단련핵감산서렬,대극륭소득산물진행HLA-A기인적제2～4외현자쌍향측서분석.결과 발현선증자적일개HLA-A*02:06:01기인피학인,이령일개HLA-A기인위신등위기인,기서렬피GenBank접수(편호위HM245348).신등위기인서렬통과IMGT/HLA 수거고BLAST,여최상근적A*02:01:01:01상비,재제3외현자상유1개핵감산적불동,즉제383위 G>C,밀마자 128 GAG→GAC,안기산유곡안산(Glu)→천문동안산(Asp).공、환자HLA-A、B、C、DQB1위점등위기인불필배.결론 해등위기인위신적HLA-A*02:251등위기인.중국인군HLA-A 위점제3외현자핵감산서렬존재다태성.
Objective To identify a novel human leukocyte antigen (HLA) allele A*02:251 and analyze the sequences in Chinese population. Methods Routine HLA-A, -B, -DRB1 high resolution genotyping for healthy Chinese donors and patients was performed with polymerase chain reaction-sequence based typing. An unknown HLA-A allele was initially detected by HLA typing in the healthy donor. Genomic DNA of the HLA-A locus in the proband was amplified, the amplified product was cloned by PMD18-T to split the two alleles, and selected clones were sequenced. Results The sequencing results showed that a normal A*02:06:01 and a novel A*02:251 variant allele were identified. The sequence of the novel allele has been submitted to GenBank (HM245348). Nucleotide sequence alignments with HLA-A allele from the IMGT/HLA Sequence Database showed that the novel A*02 variant allele differed from the closest allele A*02:01:01:01 by nt 383 G>C (codon 128 GAG>GAC) in exon 3, which resulted in one amino acid substitution of Glu>Asp. The HLA-A, B, C and DQB1 alleles of the healthy donor did not match with that of the patient. Conclusion This novel allele is officially designated as HLA-A*02:251 by World Health Organization(WHO) Nomenclature Committee (Submission ID HWS10010755). The sequence of HLA-A locus in exon 3 is confirmed to be polymorphic in Chinese population.