Lan Ke, Hu Shou-Wang, Zhang Fan, Wang Hui, Guan Wei, Ding Yu, Sun Ou-Jun, Wang Sheng-Qi
Shenzhen Yishengtang Biological Products Co. Ltd, Shenzhen 518026, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2003 Apr;11(2):174-8.
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.
HLA基因构成一个高度多态的多基因系统。在本研究中,HLA - B寡核苷酸芯片是通过使用一组来自HLA - B基因第2外显子和第3外显子多态性区域的序列特异性寡核苷酸探针制造的,这些探针通过微阵列仪点样到醛基修饰的载玻片上。此外,用作标准样品的测序HLA - B基因克隆通过PCR从第2外显子和第3外显子进行扩增。连同正确的杂交和洗涤条件,PCR产物与芯片上的阵列探针结合,杂交模式被转化为HLA - B基因型。结果表明,HLA - B寡核苷酸芯片检测标准样品的基因型与测序克隆完全相同。总之,本文介绍的用于HLA - B基因分型的寡核苷酸芯片方法是一种快速、准确、灵敏且有吸引力的高通量生化方法。