HLA-I类抗原的组织配型基因芯片的建立及应用  

DNA typing of HLA-I antigens with medium resolution method by DNA chip technique

作　　者：郭刚[1] 张瑞[1] 孙蓓[1] 张明新[1] 梁冬春[1] 

机构地区：[1]天津医科大学内分泌研究所卫生部"激素与发育重点实验室",天津300070

出　　处：《中国免疫学杂志》2007年第12期1106-1111,共6页Chinese Journal of Immunology

基　　金：天津市委重大科技攻关项目(013181811)

摘　　要：目的:应用基因芯片技术进行北方汉族人群HLA-I类抗原中分辨度分型研究,建立稳定的HLA检测芯片。方法:采用寡核苷酸芯片分型方法,依据第十三届国际组织相容性工作会议报告及相关文献,同时考虑遗传的连锁不平衡,及强脊炎、幼年型糖尿病等与HLA密切相关的遗传病等因素,选择了中国人群基因频率较高的等位基因,根据HLA-I类不同基因亚型的独特序列,完成了探针的设计与筛选,最后设计了122条探针(HLA-A56条,HLA-B66条)制成基因分型芯片。采用带荧光标记的引物,用合适的PCR方法扩增HLA-I类抗原上的多态性区域,产物与芯片上探针杂交。杂交结果经荧光扫描并用特定软件分析判断阳性探针,以此确定样品基因型。结果:所有样本的HLA-I类抗原基因分型均获成功。此中分辨度探针可分出598个I类抗原等位基因,可检出I类抗原特异性57个。结论:基因芯片用于HLA-I类抗原分型可行。其分辨率高,特异性强,可用于HLA基因分型、骨髓移植、器官移植的HLA配型、与HLA有密切关系的遗传性疾病的人群筛查。Objective:To establish a DNA typing method for HLA-I antigens with medium resolution method by DNA chip technique.Methods:The chip was made with specific medium distinguish-typing probes designed according to gene frequency of HLA-I alleles from Northern Chinese. Unsymmetrical PCR was used to amplify HLA-I exon2,3,and then the PCR products labeled and hybridized with probes on the chip.Typing of HLA-I was certified by scanning the hybridizing signals of through a set of computer software.Results:HLA-I alleles were successfully typed in 30 clinical samples .This medium-distinguishing probes were able to discern 57 HLA-I alleles accurately.Conclusion:DNA typing of HLA-I by chip has been proven to be a high-resolution and high-specific method. It is able to check out the new alleles that can not be distinguished by other methods with the same resolution., and it is more intuitional and more suitable for clinical application .

关 键 词：HLA-Ⅰ类抗原 基因芯片 中分辨度 分型 

分 类 号：R392.11[医药卫生—免疫学]