寡核苷酸芯片技术在HLA-A抗原基因分型中的应用研究

目的建立一种HLA-A位点的基因芯片分型方法,为HLA-A位点的基因分型提供一个较新的思路。方法利用基因芯片技术,根据HLA-A位点不同基因亚型的独特序列设计探针,制成分型芯片;待检测样品经PCR反应标记上荧光之后,与芯片进行杂交,根据杂交产生的荧光信号值分析确定样品HLA-A位点的基因亚型。将这一方法应用于100份标准DNA和200份无关个体的HLA-A位点基因分型并将部分样品进行测序。结果检测结果表明HLA-A基因分型芯片可准确分辨出A位点等位基因20大类,耗时2.5h。结论寡核苷酸芯片技术用于HLA-A基因分型,分辨率高、特异性强、重复性好、操作简便、结果直观,适合于法医学实践和临床应用。

Application Studies on HLA-A Genotyping by Using Microarray DNA Chip

Objective HLA-A locus is genotyped by oligoneucleotide arrays. Methods According to the sequence of HLA-A exon2 and exon3, HLA-A genotyping probes were designed and hybridized with the cyslabeled PCR products on microarrays. The signals were scanned analyzed by scanner and Imagine software. 100 standard samples and 200 unrelated Chinese Han individuals have been genotyped and some of samples have been sequenced. Results The experimental results showed that the HLA-A genotyping method was reliable and sensitive. Twenty alleles of HLA-A antigen were accurately distinguished within 2.5 h. Conclusion The DNA microarray technique for HLA-A genotyping has the characteristic of high-resolution, high-specificity and well-reproducibility which is suitable for forensic identification and clinical application.