Identification of a novel human leucocyte antigen allele B*07:110

doi:10.3969/j.issn.2095-4344.2015.01.024

Abstract

Abstract:

BACKGROUND: In recent years, with the development of China Marrow Donor Program and the improvement of human leukocyte antigen (HLA) typing technique, novel alleles of human leukocyte antigen have been discovered constantly in China.
OBJECTIVE: To identify and confirm a novel HLA allele in a Chinese individual.
METHODS: A new HLA allele was found during routine human leukocyte antigen genotyping by PCR-sequence specific oligonucleotide probes and sequencing-based typing.
RESULTS AND CONCLUSION: The HLA-B locus hybridization probe reaction patterns of this sample did not match with any known HLA-B alleles or allelic combinations. Exons 2, 3 were sequenced in both directions using HLA-B sequence primer and group-specific sequencing primer. The obtained sequence had 2nts change from B*07:02:01 at nt 226 and nt 228 where A->G (codon 76 ATA->GTG) and resulting in a coding change, 76 isoleucine (I) was changed to valine (V). This nucleotide sequence has been submitted to the GenBank nucleotide sequence database and it is available under the accession number HM989017. A novel HLA allele, HLA-B*07:110, was identified, and was named officially by the WHO Nomenclature Committee.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Tissue Engineering, HLA Antigens, Alleles, Nucleic Acid Probes

CLC Number:

R394.2

Zhou Yue, Li Jian-ping. Identification of a novel human leucocyte antigen allele B*07:110[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(1): 135-139.

Figures/Tables 2

2.1 SSO方法基因分型结果经过软件分析显示，该样本基因分型结果为HLA-A* 24，29；B* 07，44；DRB1* 07，11。但显示B*07，44 两个等位基因时出现异常提示，即17号探针假阳性(FP)，24号探针假阴性(FN)，63号探针假阴性(FN)，但反应格局显示，17号探针实际为阳性反应， 24号探针实际为阴性反应，63号探针实际为阳性反应(图1)，探针反应的整体格局清晰，背景干净。重复实验后结果发现探针分布相同，排除了实验误差，由此怀疑存在1个未认定的HLA-B新等位基因。 2.2 PCR产物双链测序结果 PCR扩增产物在第2，3外显子正反两个方向的测序结果显示该样本B基因座的序列与现有HLA数据库中已知的所有序列均不完全一致。数据分析软件显示其与同源性最高的B*44:03:01，B*07:01:02的组合在226和228位有2个核苷酸不同，2个双链的杂合点均显示为R，R表示的是A和G，但数据库中此2个核苷酸均为A，见图2，提示可能存在新等位基因。 2.3 组特异性引物测序结果为了分开双链中2个杂合的等位基因，用GSSP对原扩增产物进行再次测序反应，因GSSP仅特异性与新等位基因序列结合从而得到新等位基因的单链序列。GSSP结果显示，该样本HLA-B的1个等位基因是B*44:03:01，另一个HLA-B*07的等位基因，在所检测的第2、3外显子中，与序列同源性最高的等位基因B*07:01:02的差异是在第2外显子发生了nt 226和nt 228两个A->G核苷酸取代，导致第76位密码子由ATA->GTG，相应的导致氨基酸由异亮氨酸(I)改变为缬氨酸(V)，见图3，4。将得到的核苷酸序列的报告递交到基因数据库(GenBank)及IMGT/HLA数据库(序列号HWS10010993)，证实该新HLA等位基因为国际上首次发现，被世界卫生组织人类白细胞抗原因子命名委员会正式命名为HLA-B*07:110 (HM989017)。"

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