Bang-Ce Ye, Xiaohe Chu, Ye Fan, Songyang Li, Bincheng Yin, Peng Zuo
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
J Mol Diagn. 2005 Nov;7(5):592-9. doi: 10.1016/S1525-1578(10)60592-2.
Matching of the HLA antigens for donor-recipient in transplantation, disease predisposition or protection, population studies, and forensic testing requires accurate but simple typing methods. Here, we describe a DNA-based tissue-typing assay that determines the haplotype of the DRB1/3/4/5 loci in hy-bridization of oligonucleotide array after sample amplification. Using this multianalyte DNA hybridization system, we analyzed seven regions of exon 2 of DRB loci that have single-base discrimination. Thirty-six oligonucleotide probes complementary to the alleles of interest were immobilized on each microslide. The efficiency and specificity of identifying DRB genotypes using the oligonucleotide arrays was evaluated by blinded analysis of 147 samples from reference standards and subjects. The established method provides a rapid and inexpensive DRB "low-resolution" typing tool for prescreening a large number of samples.
在移植、疾病易感性或保护性研究、群体研究以及法医检测中，供体 - 受体的HLA抗原匹配需要准确且简便的分型方法。在此，我们描述了一种基于DNA的组织分型检测方法，该方法在样本扩增后通过寡核苷酸阵列杂交来确定DRB1/3/4/5基因座的单倍型。使用这种多分析物DNA杂交系统，我们分析了DRB基因座外显子2中具有单碱基区分的七个区域。三十六种与感兴趣的等位基因互补的寡核苷酸探针被固定在每个微载玻片上。通过对来自参考标准品和受试者的147个样本进行盲法分析，评估了使用寡核苷酸阵列鉴定DRB基因型的效率和特异性。所建立的方法为大量样本的预筛选提供了一种快速且廉价的DRB“低分辨率”分型工具。
