基因芯片快速HLA-DR52组分型

目的 用基因芯片对HLA-DR52组快速分型。方法 根据中国汉族人群常见的HLA-DRB位点及其基因多态性的独特序列，设计特异性的寡核苷酸分型探针，制成寡核苷酸芯片。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芯片的探针杂交，通过杂交产生的荧光信号确定样品的基因亚型。83份样本分别用基因分型芯片和序列特异性引物聚合酶链反应技术(PCR-SSP)对HLA-DR52组分型。结果PCR-SSP分型DR52组57个位点中，经芯片分型有2个无DR52组位点，3个PCR-SSP分型为DR52组纯合子，芯片为DR52组杂合子，1个PCR-SSP分型为非DR52组纯合子，芯片分型为含有1个DR52组位点的杂合子。结论 基因芯片能对HLA-DR52组快速、准确的分型，比PCR-SSP能更多的检出DR52组位点，特别是能将纯合子进一步分型，适合临床应用。

Rapid genotyping for HLA-DR52-associated group by oligoneucleotide arrays

Objective To develop an oligoneucleotide array for HLA-DR52 group rapid genotyping.Methods According to the special allele sequences of HLA-DRB loci in Chinese Han's population, HLA-DR52 group typing probes which were immobilized on a glass supports were synthesized. A pair of group-special primers labeled by the Cy5-dCTP were designed and were used in the PCR. The labeled PCR products with Cy5 were hybridized with array. The signals were scanned by a scanner and analyzed by Image software. 83 samples were typed by this array and the results were compared with PCR-SSP typing.Results Among 57 HLA-DR52 group loci typed by PCR-SSP,2 samples had no HLA-DR52 loci typed by array,3 DR52 group homozygotes typed by PCR-SSP were actually heterozygotes by array. The other 1 non-DR52 group homozygote identified by PCR-SSP was a heterozygote with one DR52 group locus. Conclusion The oligoneucleotide array technique is a precise, rapid molecular method for HLA-DR52 genotyping. Compared with PCR-SSP method, the genotyping chip is more sensitive and intuitionistic and suitable for clinic practice.