PCR-SSP基因分型技术检出HLAB新等位基因B~*5516一例

目的　识别确认中国人群的HLA新等位基因。方法　使用PCR SSP以及以测序为基础的分型技术 ,分析HLA新等位基因和B 5 5 0 2基因顺序的差异及血清学方法分析特异性。结果　在四川成都骨髓供者中检测出一例新的HLA B等位基因。该基因和B 5 5 0 2基因顺序的差异 ,只是在外显子 2区域中 97C >A一个碱基取代 ,导致相应的密码子 33由酪氨酸变为组氨酸。血清学分析表明B 5 5 16与B2 2特异性相关。并由此建立起PCR SSP方法鉴定B 5 5 16基因。结论　四川成都骨髓库中检出的HLA B等位基因是HLA新等位基因 ,2 0 0 3年 8月已被世界卫生组织 (WHO)命名为HLA B 5 5 16。在大约 14 0 0例随机骨髓供者中 ,未发现其他带有B 5 5 16基因的个体。

Detection of a novel allele HLA-B*5516 using PCR-SSP genotyping technique

Objective To identify HLA novel allele in Chinese population.Methods A new HLA-Ballele was initially detected by an unusual PCR-SSP reaction pattern in a potential bone marrow donor.Molecular cloning and sequencing were used to identify the difference between the new allele and HLA-B 5502 allele.Results The B 5516 allele differs from the closest matching HLA sequence of B 5502 by a single nucleotide substitution of T>C at position 97 in exon 2,resulting in an amino acid change from Tyr (TAC) to His (CAC) at codon 33. Serology study revealed that B 5516 is associated with B22 specificity. A PCR-SSP method was developed to distinguish B 5516 from other B 55 alleles.Conclusions A novel HLA-Ballele,B 5516,has been identified in a potential bone marrow donor using PCR-SSP genotyping technique.No further individuals with B 5516 were detected in 1,400 Chinese bone marrow blood donors.