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目的应用基因芯片技术进行北方汉族人群人类白细胞抗原(HLA)Ⅰ、Ⅱ类中分辨度分型研究。方法根据中国北方汉族人群HLA-Ⅰ、Ⅱ类常见的基因位点以及临床应用中对分型分辨度的实际需要,设计特异性寡核苷酸中分辨度分型探针,制成基因分型芯片。采用带荧光标记的引物,用合适的PCR方法扩增HLA-Ⅰ、Ⅱ类抗原上的多态性区域,产物与芯片上探针杂交。杂交结果经荧光扫描并用特定软件分析判断阳性探针,以此确定样品基因型。共检测30份临床样本。结果所有样本的HLA-Ⅰ、Ⅱ类抗原基因分型均获成功。此中分辨度探针可分辨出598个Ⅰ类抗原等位基因,511个Ⅱ类抗原等位基因,可捡出Ⅰ抗原特异性57个,Ⅱ抗原特异性30个。结论基因芯片用于HLA-Ⅰ、Ⅱ类抗原分型可行。
Objective To study the resolution typing of human leukocyte antigen (HLA) Ⅰ and Ⅱ in northern Han population using gene chip technique. Methods According to the common HLA-Ⅰ and Ⅱ genomic loci of Han nationality in northern China and the actual need for resolution in clinical practice, a specific oligonucleotide-based resolution typing probe was designed to generate genotyping chip. Fluorescently labeled primers were used to amplify the polymorphic regions of HLA class I and class II antigens using appropriate PCR methods and the products were hybridized to on-chip probes. Hybridization results were identified by fluorescence scanning and specific software to determine positive probes to determine sample genotype. A total of 30 clinical samples were tested. Results All samples were successfully genotyped HLA class I and class II antigens. The resolution probe can distinguish 598 class Ⅰ antigen alleles and 511 class Ⅱ antigen alleles, which can detect 57 specific I antigen and 30 specific antigen Ⅱ. Conclusion Gene chips for HLA-Ⅰ, Ⅱ antigen type feasible.