DNA微阵列技术检测HLA-DRB1基因分型  

Detection for HLA-DRB1 genotyping by DNA microarray

作　　者：周转[1] 胡守旺[2] 张帆[2] 

机构地区：[1]河南省焦作市人民医院,河南焦作454002 [2]中山大学达安基因诊断中心,广东广州510665

出　　处：《分子诊断与治疗杂志》2009年第3期156-160,共5页Journal of Molecular Diagnostics and Therapy

基　　金：国家863计划重大专项(2002AA2Z2032)

摘　　要：目的采用DNA微阵列分型方法,对HLA-DRB1基因进行分型研究。方法设计分型检测寡核苷酸探针,制备HLA-DRB1基因分型微阵列。设计PCR引物,以1:20引物比例不对称扩增HLA-DRB1基因的第2外显子,Cy3荧光标记扩增产物,作为杂交模板。用差异选择法杂交信号强且特异性好的探针。Cy3荧光标记PCR扩增产物与微阵列探针杂交反应,结果经荧光扫描,并用分型软件分析判断阳性探针和HLA-DRB1基因型。结果10例临床血样的DNA微阵列分型结果与PCR-SSP分型结果相符,20例未知血样的DNA微阵列分型结果与DNA测序分型结果符合率为97%。结论DNA微阵列技术具有高通量、简便、低成本的优势,采用DNA微阵列分型方法,对HLA-DRB1基因进行分型研究,表明HLA-DRB1DNA微阵列分型为一种可行的基因分型新技术。Objective To develop a DNA microarray method for HLA-DRB1 typing.Methods The specific oligonucleotide probes for HLA-DRB1 genotyping was designed and spotted on glass slides using a microarray spotter.DNA sample was unsymmtric PCR amplified for the exon 2 of HLA-DRB1 and was labeled with the fluorescent dye Cy3.Following hybridization between the Cy3-labeled PCR products and the oligonucleotide probes of each microarray on the slides and wash,the amount of probe hybridizing to each DNA sample was measured with a microarray scanner and the image was analyzed with a computer software,which was used to identify the positive probes and to determine the HLA-DRB1 genotype of each sample.Results There was good concordance between the genotyping results by oligonucleotide microarray and those of PCR-SSP or SBT.Conclusion The procedure described here has the potential of providing a rapid,simple,economical and accurate way for HLA-DRB1 genotyping.

关 键 词：HLA-DRB1 基因分型 DNA微阵列 PCR-SSP 

分 类 号：R392.1[医药卫生—免疫学]