寡核苷酸芯片用于HLA-B分型的初步研究  

Preliminary Study on HLA-B Genotyping by Oligonucleotide Chips

作　　者：蓝柯 胡守旺 张帆 王晖 管伟[2] 丁雨 孙偶军[2] 王升启[2] 

机构地区：[1]深圳益生堂生物企业有限公司,深圳518026 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《中国实验血液学杂志》2003年第2期174-178,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金重点资助项目编号3 98890 0 1

摘　　要：HLA基因具有复杂的多态性。本研究根据HLA B基因序列的多态性 ,设计了一套寡核苷酸探针 ,并用基因芯片点样仪点样到醛基修饰的载玻片上 ,制成寡核苷酸芯片用于HLA B基因的分型。本研究以克隆并已测序的HLA B基因序列作为标准样品 ,经PCR分别扩增其具有多态性的第二和第三外显子 ,PCR扩增产物与芯片进行杂交 ,利用分析软件将杂交模式转变成为HLA B基因型。结果表明 ,利用芯片进行分型的结果与标准样品序列结果完全符合。结论 :寡核苷酸芯片用于HLA B基因分型是一种简便。HLA genes constitute a highly polymorphic multigene system. In the present study, HLA B oligonucleotide chips were manufactured by using a set of sequence specific oligonucleotide probes derived from polymorphic regions in exon 2 and exon 3 of HLA B gene spotted by microarrayer onto the aldehyde modified glass slides. In addition, the sequenced HLA B gene clones used as standard samples were amplified from exon 2 and exon 3 by PCR. Together with the correct hybridization and wash conditions, the PCR products were bound with the array probes on the chip, and the hybridization patterns were transformed to HLA B genotypes. The results showed that the genotypes of standard samples by the HLA B oligonucleotide chips were completely identical with the sequenced clones. In conclusion, the oligonucleotide chip method presented here for HLA B genotyping is a rapid, accurate, sensitive and attractive high throughput biochemical way.

关 键 词：寡核苷酸芯片 HLA-B分型 多态性 白细胞抗原 血清学 

分 类 号：R446.6[医药卫生—诊断学]