DNA芯片技术检测HLA-DR1,DR51组基因型  

Genotyping of HLA-DR1,DR51-associated group by DNA microarray

作　　者：肖家全[1] 李成涛 谭建明[1] 康敏华 李瑶 丁言德[1] 沈瑾[1] 

机构地区：[1]上海市第一人民医院上海市器官移植临床学中心,现在浙江省人民医院200080 [2]上海联合基因集团博星芯片研究所

出　　处：《中华泌尿外科杂志》2003年第11期735-737,共3页Chinese Journal of Urology

基　　金：上海市科学技术发展基金资助项目 ( 0 2 49190 0 5 ) ;全军"十五"重大课题"杰出人才"基金资助项目 ( 0 1J0 0 3 )

摘　　要：目的　采用DNA芯片技术对HLA DR1,DR5 1组基因分型。　方法　根据编码HLA DR1,DR5 1组抗原等位基因序列设计特异分型探针 ,制作DNA分型芯片。通过组间特异引物扩增基因组相应区段DNA ,扩增中用Cy5 dCTP荧光标记 ,扩增标记后的产物与芯片探针杂交 ,通过杂交产生的荧光信号及分布格局确定样品基因亚型。分型结果用标准DNA和PCR SSO验证。　结果　130份样本共检出HLA DR1,DR5 1组位点 34个 ,包括 18个DR15 ,8个DR16 ,6个DR10 ,2个DR1,无假阳性或假阴性 ,准确率和重复率均达 10 0 %。检测总耗时约 3.5h。　结论　DNA芯片用于HLA DR1、DR5 1组基因分型具有高分辨度、高特异性和简单快速的优点 ,适于器官移植临床应用。Objective To develop a DNA microarray for HLA-DR1,DR51 group genotyping. Methods According to the specific allelic sequences coding HLA-DR1,DR51 loci,HLA- DR1,DR51 group typing probes which were immobilized on a glass support were synthesized.A pair of group-specific primers labeled by the Cy5-dCTP were designed,then the primers were used in the PCR,thus the PCR products were labeled with Cy5.The labeled PCR products were hybridized with array.The signals were scanned by scanner and analyzed by image software.The typing results were confirmed by standard DNA and PCR-SSO. Results A total of 130 samples were typed by this DNA array.There were 34 HLA-DR1,DR51 group loci typed by DNA array.Among them,18 loci were DR15,8 were DR16,6 were DR10 and 2 were DR1.No false positive or false negative typing results occurred.The accuracy and reproducibility were 100% and the overall time of genotyping was about 3.5 hours. Conclusions DNA array technique is a precise,rapid molecular method of high resolution power and high specificity for HLA-DR1,DR51 genotyping,which is applicable to clinical transplant practice.

关 键 词：DNA芯片技术 检测 HLA—DR1 DR51组基因型 人白细胞抗原 

分 类 号：R346[医药卫生—基础医学]