应用毛细管参考链介导的构象分析法进行HLA-DR4等位基因分型  

HLA-DR4 allele assignments by reference strand mediated conformation analysis using capillary-based genetic analyzer

作　　者：毕颎[1] 陈规划[2] 刘泽寰[3] 黄丽英[4] 徐安龙[3] 

机构地区：[1]中山大学附属第一医院外科实验室 [2]中山大学附属第三医院肝移植科,广州510630 [3]中山大学生命科学院,广州510275 [4]中山大学达安基因诊断中心,广州510080

出　　处：《免疫学杂志》2005年第2期145-147,共3页Immunological Journal

摘　　要：目的　应用毛细管参考链构象分析法对HLA DR4进行等位基因分型,并与测序分型结果相比较,评价该方法的准确性及实用性。方法　首先应用FAM标记的引物扩增HLA DRB1等位基因为纯合型的样本DRB1 0 80 3、DRB1 0 90 1的第二外显子作为参考链,与标准样品HLA DRB1第二外显子杂交获得异源双链,借助测序仪通过毛细管电泳建立中国人HLA DR4 8个高频率等位基因的标准迁移率,再对30例DR4样本进行分型,并与测序分型法检测结果进行比较。结果　30例样本中2 8例与测序法测得的等位基因结果一致。结论　毛细管参考链介导的构象分析法具有准确性高、分辨率高的优势,与测序法相比具有成本低、高通量的特点,值得在移植、相关疾病等领域的HLA分型中推广应用。Objective To evaluate the accuracy and application of reference strand mediated conformation analysis (RSCA) using ca- pillary -based genetic analyzer in HLA-DR4 allele assignments, by comparison with sequence based typing (SBT). Methods FAM labelled PCR products from homozygote DRB1*0803 and DRB1*0901 were the fluorescently labeled references. Heteroduplexes were generated by HLA-DRB1 exon 2 hybridization of reference and standard sample. The standard mobility of 8 alleles with high frequency in Chinese was established through RSCA using capillary-based genetic analyzer. A total of 30 samples were typed by RSCA in contrast with SBT. Results Twenty eight of 30 samples typed by capillary RSCA and SBT showed complete concordance in allece assignments. Conclusion Capillary RSCA is useful in HLA typing with high accuracy, high resolution, high throughput, but low cost.It is desirable to be applied in HLA assignment for transplantation and diseases related research.

关 键 词：参考链介导的构象 等位基因 HLA—DR4分型 

分 类 号：R346[医药卫生—基础医学]