HLA-DRB1基因芯片与PCR-SSP分型的比较  

Comparative Study of HLA-DRB1 Genotyping by Oligoneucleotide Arrays and PCR-SSP

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

机构地区：[1]上海市第一人民医院-上海市器官移植研究中心,上海200080 [2]联合基因集团博星芯片研究所,上海200092

出　　处：《复旦学报（医学版）》2003年第1期68-70,共3页Fudan University Journal of Medical Sciences

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"杰出医学人才基金资助项目 ( 0 1J0 0 3)

摘　　要：目的　比较基因芯片和PCR SSP用于汉族南方人群HLA DRB1基因分型的结果 ,评价分型芯片的准确性。方法　 77份待分型样本 ,分别用等位基因特异的PCR SSP和基因分型芯片对DRB1分型。芯片分型通过组间特异引物扩增基因组DNA ,扩增中用荧光标记。扩增标记后的产物与分型芯片的探针杂交 ,通过杂交产生的荧光信号确定样品的HLA DRB1基因亚型 ,比较两种方法分型所获得的结果 ,不吻合的样本全部经第三方验证或测序。结果　 77份样本 ,两种方法分型全部成功。分型结果的吻合率为 93 %。不吻合样本 6份 ,其中SSP定型纯合子 4份 ,芯片分型全部为杂合子。经第三方验证 ,证实芯片对纯合子和杂合子的分型全部正确。另外 2份不吻合的样本经测序 ,证实SSP分型错误 1份 ,芯片分型错误 1份。芯片的重复率为 96%。结论　基因芯片是一种理想的分型方法 ,具有特异性高、重复性好、操作简便、所需样本量少。Purpose: To evaluate the accuracy and reliability of oligoneucleotide array in comparison with polymerase chain reaction with sequence-specific primers (PCR-SSP) in identification of human leukocyte antigen-DRB1 (HLA-DRB1) alleles in Southern Han's population. Methods: A total of 77 samples were enrolled in the study. HLA-DRB1 genotyping were performed by PCR-SSP and oligoneucleotide array. A pair of group-specific primers were designed according to the sequence of HLA-DRB1 exon2, then the primers and the Cy5-dCTP were used in the PCR, thus the PCR products were labeled with Cy5. The labeled PCR products were hybridized with the specific allele typing probes which were immobilized on a glass support, and the signals were scanned by scanner and then analyzed. The samples which the HLA-DRB1 typing results by arrays and PCR-SSP were discordant were verified by PCR-SSOP or sequencing. Results: All the samples have been genotyped by HLA array and PCR-SSP successfully. 4 homozygous samples typed by PCR-SSP were actually heterozygous by array. The other 2 unidentified samples were typed by sequencing, the results showed that PCR-SSP made a mistake for 1 samples and array for 1. Conclusions: The oligoneucleotide array technique is a precise, rapid molecular method for HLA-DRB1 genotyping, and has the advantage of high specificity and integration.

关 键 词：HLA-DRB1 基因芯片 PCR-SSP 器官移植 排斥反应 基因分型 

分 类 号：R392.4[医药卫生—免疫学] Q78[医药卫生—基础医学]