Zhang Xu, Jiang Shu-yi, Li Xiao-feng, Zhang Kun-lian, Chen Yang, Liu Xian-zhi, Li Jian-ping
Liaoning Blood Center, Shenyang, Liaoning 110044, P. R. China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2012 Feb;29(1):91-4. doi: 10.3760/cma.j.issn.1003-9406.2012.01.022.
To identify a novel human leukocyte antigen(HLA) allele in Chinese population.
HLA typing was carried out with polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP). The HLA-B exons 1-7 of the proband were amplified and the product was cloned using a TOPO TA cloning sequencing kit to separate the two alleles. Both strands of exons 2 and 3 of selected colonies were sequenced. Sequence-based typing (SBT) was used to identify and analyze the difference between the new allele and the closest matching HLA-B allele.
HLA typing indicated a SSOP pattern which did not match with known HLA-B alleles. The results of the sequencing suggested the HLA-B alleles of the proband as B59:01 and a novel allele. The HLA-B exon 3 sequence of the novel allele was different from any known alleles. This allele differs from the closest matching B54:06 allele by 6 nucleotides, which included nt486 (G to C), nt527 (A to T), nt538 (T to C), nt539 (G to T), nt559 (C to A) and nt560 (T to C) in exon 3, resulting in substitutions of three amino acids including Glu to Val at codon 152, Trp to Leu at codon 156 and Leu to Thr at codon 163.
A novel HLA-B allele has been identified and has been designated as HLA-B*54:09 by WHO Nomenclature Committee for Factors of the HLA System.
在中国人群中鉴定一种新的人类白细胞抗原（HLA）等位基因。
采用聚合酶链反应-序列特异性寡核苷酸探针（PCR-SSOP）进行HLA分型。对先证者的HLA-B外显子1-7进行扩增，并使用TOPO TA克隆测序试剂盒对产物进行克隆以分离两个等位基因。对选定菌落的外显子2和3的两条链进行测序。基于序列的分型（SBT）用于鉴定和分析新等位基因与最匹配的HLA-B等位基因之间的差异。
HLA分型显示出一种与已知HLA-B等位基因不匹配的SSOP模式。测序结果表明先证者的HLA-B等位基因为B59:01和一个新的等位基因。新等位基因的HLA-B外显子3序列与任何已知等位基因都不同。该等位基因与最匹配的B54:06等位基因在第3外显子中有6个核苷酸不同，包括nt486（G到C）、nt527（A到T）、nt538（T到C）、nt539（G到T）、nt559（C到A）和nt560（T到C），导致三个氨基酸发生替换，包括第152密码子处的Glu替换为Val、第156密码子处的Trp替换为Leu以及第163密码子处的Leu替换为Thr。
已鉴定出一种新的HLA-B等位基因，世界卫生组织HLA系统因子命名委员会已将其命名为HLA-B*54:09。
