论文部分内容阅读
目的建立HLA-A位点等位基因的PCR-SBT高分辨分型方法,探讨DNA测序技术在脐血库样本HLA分型中的应用价值。方法利用PCR产物直接测序,对广州脐血库保存的547份脐血样本进行HLA-A位点2、3、4外显子的序列分析,由分型结果得出基因频率,与中华(上海)骨髓库北方人群、上海地区人群及德国白种人进行比较。结果采用PCR-SBT分型方法并结合分析软件确定了全部样本的HLA-A基因型,广州地区人群HLA-A等位基因以A*110101(30.8%)最为常见,其后依次是A*24020101/02L(16.18%)、A*0207(11.88%)、A*3303(9.42%)。A*110101在广州汉族人群中出现的频率明显高于中华(上海)骨髓库北方人群,而A*010101、A*3001明显低于后者;在HLA-A2亚型人群中,A*020101在广州、上海两地汉族人群中的频率明显低于德国白种人,而广州汉族人群中A*020101与A*0206均明显低于上海汉族人,但A*0203明显高于后者。结论基于核酸序列测定的HLA分型技术能够直接、准确、快速地进行高分辨分型,将有助提高无亲缘关系供者脐血移植的临床效果。改进实验条件、升级分型软件,可以降低试剂成本和节约时间。
Objective To establish a high-resolution PCR-SBT method for HLA-A locus alleles and to explore the value of DNA sequencing in the HLA typing of cord blood samples. Methods The sequence of exon 2, 3 and 4 of HLA-A locus was analyzed by direct sequencing of PCR products in 547 cord blood samples stored in Guangzhou cord blood bank. Bone marrow pool north of the population, the Shanghai area population and German white compared. Results The HLA-A genotypes of all the samples were determined by PCR-SBT typing and analysis software. The HLA-A allele was most common in A * 110101 (30.8%) in Guangzhou, followed by A * 24020101 /02L(16.18%),A0207(11.88%),A3303(9.42%). The frequency of A * 110101 in Han population in Guangzhou was significantly higher than that in North China (Shanghai) Marrow Bank, but A * 020101 and A * 3001 were significantly lower in A * The frequencies of Han population in Guangzhou and Shanghai were significantly lower than those in German white people. However, A * 020101 and A * 0206 in Guangzhou Han population were significantly lower than those in Shanghai Han, but A * 0203 was significantly higher than that in Han. Conclusion The HLA typing technique based on nucleic acid sequencing can directly, accurately and rapidly classify high-resolution genotypes, which will help to improve the clinical effect of unrelated donor cord blood transplantation. Improve experimental conditions, upgrade typing software, reagents can reduce costs and save time.