人类白细胞抗原-A基因芯片分型研究

目的 探索人类白细胞抗原-A（HLA-A）基因芯片分型，为器官移植临床配型服务。方法 根据中国汉族南方人常见的HLA-A位点基因及其多态性的独特序列，设计并合成48条特异性的寡核苷酸分型探针，将其点在玻片上，制成芯片。基因组DNA通过组间特异性引物扩增，并用荧光素Cy5标记。标记后的产物与结合在芯片上的探针进行杂交，通过荧光扫描仪获得杂交产生的荧光信号值，再经过计算机软件自动分析，确定样品的HLA-A基因亚型。用该方法对120份样本进行HLA-A基因分型。结果 120份待检样本，其中6份因PER无产物，不能分型。1份信号杂乱，不能分型。其余113份样本分型成功。实际检出A抗原特异性结果为A2（含A203）：56；A11（含A1101）：52；A24：33；Al：8；A30（含A3001）：7；A33：21；A26：1；A29：2；A31：3；A68：2；A3：9；A32：1。未检出A＊3002基因型。整个检测耗时约4．5h。芯片检测的重复率为100％。结论 HLA-A基因芯片是一种理想的分型方法，具有特异性高、重复性好、操作简便、所需样本量少、结果判读容易、一次可作多份样本的优点，适合临床器官移植配型应用。

Human leucocyte antigen A genotype by oligonucleotide array

Objective To develop an oligonucleotide array for HLA-A genotype and to use it for transplantation tissue match. Methods According to the special allele sequences of HLA-A loci in Southern China Han's population, 48 typing probes which were immobilized on a glass support were synthesized. Three group-special primers were designed according to the sequence of HLA-A exon2 and exon3, then the primers and the Cy5-dCTP were used in the PCR, thus the PCR products were labeled with Cy5. The labeled PCR products were hybridized with array, and the signals were scanned by scanner and then analyzed by image software. One hundred and twenty samples were differentiated by this array. Results All the samples except 6 without PCR products and one contaminated were identified by HLA array. The results of HLA-A antigen specificity were as follows: A2(including A203) : 56; A11(including A1101): 52; A24: 33; A1: 8; A30(including A3001): 7; A33: 21; A26: 1; A29: 2; A31: 3; A68: 2; A3: 9; A32: 1. No A * 3002 had been founded. The overall time for whole processing was 4.5 hours and the reproducibility was 100% . Conclusion HLA-A oligonucleotide array technique is a precise, intuitionistic and fast molecular technique for HLA-A genotyping and is suitable for transplant histocom-patibility genotyping.