论文部分内容阅读
目的 用基因芯片对HLA DR5 2组快速分型。方法 根据中国汉族人群常见的HLA DRB位点及其基因多态性的独特序列 ,设计特异性的寡核苷酸分型探针 ,制成寡核苷酸芯片。通过组间特异引物扩增基因组DNA ,扩增中用荧光标记 ,扩增标记后的产物与芯片的探针杂交 ,通过杂交产生的荧光信号确定样品的基因亚型。 83份样本分别用基因分型芯片和序列特异性引物聚合酶链反应技术 (PCR SSP)对HLA DR5 2组分型。结果 PCR SSP分型DR5 2组 5 7个位点中 ,经芯片分型有 2个无DR5 2组位点 ,3个PCR SSP分型为DR5 2组纯合子 ,芯片为DR5 2组杂合子 ,1个PCR SSP分型为非DR5 2组纯合子 ,芯片分型为含有 1个DR5 2组位点的杂合子。结论 基因芯片能对HLA DR5 2组快速、准确的分型 ,比PCR SSP能更多的检出DR5 2组位点 ,特别是能将纯合子进一步分型 ,适合临床应用。
Objective To genotype HLA DR5 2 by gene chip. Methods Based on the unique HLA DRB loci and their unique genetic polymorphisms in Chinese Han population, oligonucleotide probes were designed to generate oligonucleotide microarray. Genomic DNA was amplified by specific primers between groups. Fluorescent labeling was used to amplify the labeled product. The amplified product was then hybridized with the probe of the chip. The fluorescence signal generated by hybridization was used to determine the genotype of the sample. 83 samples were genotyped by chip and sequence-specific primer polymerase chain reaction (PCR SSP) HLA DR5 2 component type. Results Among 5 7 sites of PCR SSP genotyping, there were 2 DR5 2 loci on the chip, 3 PCR SSPs were classified as DR5 2 homozygotes and DR5 2 heterozygotes, One PCR SSP genotyped to a non-DR5 2 homozygote and a chip-typed heterozygote containing one DR5 2 locus. Conclusion Gene chip can rapidly and accurately classify HLA DR5 2 group, and can detect more DR5 2 loci than PCR SSP. In particular, it can further homogenize the homozygotes and is suitable for clinical application.