HLA-DR位点的基因芯片分型与临床应用

HLA-DR genotyping and clinical application using Microarray DNA Chip

目的 :建立一种新型的HLA DR位点的基因分型方法 ,为HLA DR的基因分型提供一个较新的思路。方法 :利用基因芯片技术 ,根据HLA不同基因亚型的独特序列设计探针 ,制成分型芯片 ;待检测样品经PCR反应标记上荧光之后 ,与芯片进行杂交 ,根据杂交产生的荧光信号值分析确定样品DR位点的基因亚型。将这一方法应用于 70份标准DNA和 2 0 0份医院移植供受者的HLA DR基因分型并将部分样品进行基因测序。结果 :检测结果表明HLA DR基因分型芯片可准确分辨出DR位点等位基因 30大类 ,耗时 3h。结论 :基因芯片用于HLA DR的基因分型 ,分辨率高、特异性强、重复性好、操作简便 ,对比常规的PCR SSP和PCR SSO方法 ,HLA DR基因芯片方法更为直观 ,并具有集成化优势 ,可以在一张芯片上同时检测HLAI类的A、B位点 ,并实现一张芯片多人份 ,适合于临床应用和骨髓库、脐血库的建立。

Objective:To establish a new method performed on an DNA chip for genotyping HLA DR and supply a new view.Methods:According to the particular sequence of HLA DR exon2,HLA DR genotyping Chip was made,then the labeled PCR products hybridized with them,the signals were sanned by sanner and analyzed by Imagene software.70 standard DNA and 200 donor recipients have been genotyped and some of samples have been sequenced.Results:The experimental results showed that the HLA DR genotyping chip made are accurate and sensitive.Thirty alleles of HLA DR were accurately distinguished and its overall time of DNA typing was 3 hours.Conclusion:This proved that the DNA Microarray technique is good for DR genotyping and high resolution,high specificity,well reproducibility.Compared with PCR SSP and PCR SSO methods,the genotyping chip method is more intuitionistic and has the advantage of integration.It can also genotype HLA A,B alleles and many persons in a chip at the same time.It is more suitable for clinical application and establishment of marrow bank and umbilical cord bank.