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.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
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)
