应用反向斑点杂交技术进行脐血HLA-DRB基因分型方法的建立  

Establishment of A Method for HLA-DRB Genotyping in Cord Blood by Reverse Dot-Blot Hybridization Technique

作　　者：黄以宁[1] 廖灿[1] 汤雪薇[1] 李焱[1] 谢杏梅[1] 曾瑞萍[2] 

机构地区：[1]广州市妇婴医院,广州市脐血库广州510180 [2]中山医科大学医学遗传教研室,广州510089

出　　处：《中国实验血液学杂志》2002年第2期148-152,共5页Journal of Experimental Hematology

摘　　要：HLA分子的多态性对于移植有十分重要的意义。常用的HLA基因分型方法中SSOP基因分型方法有较高分辨率 ,但操作过程复杂 ,仅适于大样本的HLA分型。PCR SSP基因分型方法分辨率较低 ,但操作过程简单 ,适于临床要求。有必要建立一种简便、快速、价廉、高分辨率的基因分型方法。本研究取 6 3份已知HLA DRB型的脐血标本 ,经盐酸胍方法提取DNA用于反向斑点杂交 (RDB)基因分型 ,同时采用SSOP和PCR SSP方法进行比较。结果表明 ,所有样本用RDB方法基因分型均获得成功 ,可准确地分辨 6 0个HLA DRB等位基因 ,且结果与用SSOP和PCR SSP方法的结果完全一致。结论提示 ,RDB方法可准确地分辨HLA DRB等位基因 ,分辨率高 ,操作简单 。The HLA system was discovered by virtue of the fact that it was polymorphic. The impetus for its discovery was the search for polymorphic antigens to match for transplantation, by analogy with the human red cell blood groups. The most usually DNA method of HLA typing is sequence specific oligonucleotides(SSO) and PCR sequence specific primers(SSP). SSO technique is perfectly suited for analyzing large numbers of samples, it is not suitable for individual or small numbers. The SSP method is ideal for typing individual samples, but it is costly and requires high capacity thermal cycles for larger numbers of samples. To set up a simple, quick, cheap and high resolution DNA method, were collected sixty three cord blood samples from Guangzhou Cord Blood Bank, got DNA from blood by the traditional guanidine hydrochloride distillation method. Each sample was simultaneously typed by SSOP, PCR SSP and reverse dot blot hybridization(RDB) methods. All of typed is success. The results of three DNA methods are consistent each other. 60 HLA DRB1 allellas could be accurately distingushed with the RDB method. Our results show that RDB method is a simple, quick, cheap and high resolution method for HLA DRB types. It can be used in any HLA typing.

关 键 词：反向斑点杂交技术 HLA-DRB基因分型 脐血 特异性序列引物PCR 序列特异性寡核苷酸探针 白细胞抗原 

分 类 号：R446.62[医药卫生—诊断学] Q503[医药卫生—临床医学]