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目的 应用寡核苷酸芯片分型方法 ,对HLA A基因进行分型研究。方法 采用不对称PCR方法 ,扩增HLA A基因的第 2 ,3外显子 ,荧光标记扩增产物 ,作为杂交模板。设计分型检测寡核苷酸探针 ,制备HLA A基因分型检测芯片。采取差异选择法 ,筛选强杂交信号和高特异性的分型探针。探索了探针长度、探针位置等对杂交信号的影响。杂交结果经荧光扫描 ,并用分型软件分析判断阳性探针和HLA A基因型。结果 30例临床样本芯片分型结果与PCR SSP及DNA测序分型结果相符。结论 采用寡核苷酸芯片技术对HL A基因分型是种好的方法 ,具有测速快、成本低、高通量的优点。
Objective To study the typing of HLA A gene by oligonucleotide microarray. Methods The asymmetric PCR method was used to amplify the exons 2 and 3 of HLA A gene and amplify the fluorescently labeled products as a hybridization template. Designing a typing detection oligonucleotide probe to prepare HLA A genotyping detection chip. Differential selection method was used to screen strong hybridization signals and high specificity typing probes. The effect of probe length and probe position on the hybridization signal was explored. Hybridization results were fluorescently scanned and genotyping software was used to analyze positive probes and HLA A genotypes. Results The typing results of 30 clinical samples were consistent with those of PCR SSP and DNA sequencing. Conclusion The oligonucleotide microarray technique is a good method for genotyping HL A, which has the advantages of fast speed, low cost and high throughput.