摘要
目的识别确认中国汉族人群中的HLA新等住基因。方法使用FLOW—SSO对骨髓库样本进行常规分型，发现一份样本DRB1位点的分型结果为DRB1＊09，1109。用PCR．SBT方法对该样本进行确认，发现在外显子2有1个位置与数据库不相符。用组特异性引物分别扩增DRB1＊09及DRB1＊11基因，对外显子2进行双向测序，发现一个与DRB1＊1128序列相近的新等位基因。分析该等位基因与DRB1＊1128序列的差异。用EB病毒感染外周血B淋巴细胞，建立该等位基因的无限增殖化B淋巴细胞系。结果该等住基因与DRB1＊1128相比在外显子2有1个碱基的改变，碱基189A→G，导致密码子34Q（CAA）→Q（CAG），该氨基酸是同义突变。成功建立了该等住基因的无限增殖化B淋巴细胞系。结论该等位基因为新的HLA—DRB1等位基因，该基因序列已提交Genbank，注册号为FJ870104，已于2009年4月被世界卫生组织HLA因子命名委员会正式命名为HLA—DRB1＊112802（上报编号HWS10006282）。
Objective To identify HLA novel allele in Chinese Han individual. Methods Initially,a sample from CMDP （Chinese Marrow Donor Program） was typed by FLOW-SSO （sequence-specific oligonucleotide） and its genotype was DRB1 ＊ 09xx, 1109. But DRB1 ＊ 1109 was a lower frequency allele. Heterozygous sequence-based typing （SBT） showed there was one difference compared with database in exon2. To separate the two alleles and to determine which allele was novel ,amplified the HLA-DRB1 ＊ 09 and HLA-DRB1 ＊ 11 alleles separately by using a commercial kit for the single allele- specifie sequencing strategy. To prepare B-lymphoblastoid cell lines of the novel HLA allele by using Epstein-Barr virus infected Bqymphoblastoid cells in the peripheral blood. Results After sequence-based typing,two different alleles were assigned： DRB1 ＊ 090102 and a second allele its sequences were identical to DRB1 ＊ 1109 except for a single nueleotide substitution at ntl81 where C〉T,codon32 H（CAT）〉Y （TAT）,or was identical to those of HLA-DRB1 ＊ 1128 except for one nucleotide change at nt189 （A→G）,codon34 Q （CAA） →Q （CAG）,no coding change. Immortalized B-lym- phoblastoid cell lines of the novel HLA-DRB1 ＊ 11 allele were achieved. Conclusion The sequence is submitted to Gen- bank and the accession number is FJ870104. The allele has been officially named HLA-DRDB1 ＊ 112802 by the WHO Nomenclature committee in April 2009（HWS10006282）.
出处
《现代检验医学杂志》
CAS
2009年第4期9-12,共4页
Journal of Modern Laboratory Medicine
基金
基金项目：2009年深圳市科技计划项目（资助项目2009-02120号）
