Simultaneous genotyping of DRB1/3/4/5 loci by oligonucleotide microarray
Simultaneous genotyping of DRB1/3/4/5 loci by oligonucleotide microarray
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DOI：
10.1016/s1525-1578(10)60592-2
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发表时间：
2005-11-01
影响因子：
4.1
通讯作者：
Zuo, P
中科院分区：
文献类型：
--
作者：
Ye, BC;Chu, XH;Zuo, P
在移植、疾病易感性或保护、人群研究和法医学检测中，供受者的HLA抗原匹配需要准确但简单的分型方法。在这里，我们描述了一种基于DNA的组织分型方法，该方法在样品扩增后的寡核苷酸阵列杂交中确定DRB 1/3/4/5位点的单倍型。使用这种多分析物DNA杂交系统，我们分析了7个区域的DRB基因座的外显子2具有单碱基歧视。将与感兴趣的等位基因互补的36个寡核苷酸探针固定在每个载玻片上。通过对来自参考标准和受试者的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。建立的方法提供了一个快速和廉价的DRB“低分辨率”分型工具，用于预筛选大量的样品。
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 hybridization 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.