寡核苷酸DNA Microarray用于HLA DRB1基因分型的研究

对寡核苷酸DNA Microarray用于HLA DRB1基因分型的技术进行研究。常规的酚/氯仿法提取标准血样基因组DNA，在DRB1的exon2区域设计一对引物，经PCR扩增基因组相应区段并用Cy5-dCTP进行标记。设计寡核苷酸分型探针，将探针固定在APS-PDC法制作的DNA Microarray上，用标记的PCR产物与之杂交，扫描仪对杂交效果进行扫描，Imagene软件对杂交图像进行分析。共检测了33例标准血样的HLA DRB1基因型。检测结果证明研制的DNA Microarray准确、灵敏。DNA Microarray技术可以有效地检测DRB1等位基因，对比常规的PCR-SSP和PCR-SSO方法、分型基因芯片方法更为直观，并有集成化优势。

Investigation of Genotyping HLA DRB1 Gene Using Oligoneucleotide Arrays

We have developed a method performed on an oligoneucleotide array for genotyping HLA DRB1 . The routine method Phenol chloroform was used to extract genome DNA of standard samples. A pair of sense and antisense primers were designed according to the sequence of DRB1 exon2 ,then the primers and the Cy5 dCTP were used in the following PCR,thus the PCR products were labelled with Cy5. Many genotyping probes which were immobilized on the DNA Microarray made by APS PDC method were designed. The labelled PCR products were hybridized with them,the signals were sanned by sanner and analyzed by Imagene software. We have genotyped 33 standard samples which have 12 DRB1 subtypes. The experimental results showed that the arrays we made and the method we used are accurate and sensitive. This proved that the DNA Microarray technique is good for DRB1 genotyping. Compared with PCR SSP and PCR SSO methods,the genotyping chip method is more intuitionistic and has the advantage of integration.