一种以测序为基础的基因分型方法的建立  

A Sequence-based Typing Method for HLA-DPB1 Genotyping

作　　者：刘泽寰[1] 林蒋海[1] 符志彦[1] 潘德京[1] 徐安龙[1] 

机构地区：[1]中山大学生命科学学院,广东广州510275

出　　处：《中山大学学报（自然科学版）》2001年第5期83-87,共5页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家杰出青年基金资助项目(39725007)

摘　　要：设计了一种自动化的基因分型方法，能够直接利用ＰＣＲ产物的ＤＮＡ测序结果作为数据，通过自动化基因分型程序（ａｕｔｏｍａｔｉｃ ｇｅｎｏｔｙｐｉｎｇ ｐｒｏｇｒａｍ）与一个含有其等位基因序列的数据库进行比较，来完成高精度的基因分型工作．用这种方法对５０个随机挑选的中冈南方人的ＨＬＡ－ＤＰＢ１基因进行广基因分型，证明该方法是可行的，并从中发现了２个不明确杂合子以及一个新的等位基因序列．随后对这些样品克隆测序，发现除了２个不明确杂合子得到了进一步的区分外，其它样品都与前面的分型结果一致，显示了９６％的精确性．通过以上的研究表明，成功建立了一种高精度的、基于序列的分型（ｓｅｑｕｅｎｃｅ－ｂａｓｅｄ　ｔｙｐｉｎｇ）方法．既然该方法能成功分型ＨＬＡ－ＤＰＢ１基因，预示了该方法同样能应用于其它基因的分型．该ＡＧＰ程序可以在ｈｔｔｐ：／ｇｅｎｏｍｅ．ｚｓｕ．ｅｄｕ．ｃｎ免费下载．An automatic sequence-based typing (SBT) method is established for genotyping. It is a high-resolution genotyping method by utilizing the automatic genotyping program (AGP) to compare the DNA sequence data ob- tained by direct sequencing of PCR products with a database containing kuown allelic sequences. It offers SBT the ability to distinguish heterozygous individuals as well as identifying homozygous individuals. To validate this typing system, 50 unrelated southern Chinese individuals were typed in the HLA-DPB1 locus as an example. Two ambigu-ous heterozygotes and one novel allelic sequence were found in these individuals. The typing results were confirmed by cloning and subsequent sequencing of the cloned DNA without any discordance except two ambiguous heterozy-gotes, indicated a 96% concordance. Thus, an efficient and accurate SBT method was established by using AGP program. It was proved applicable in genotyping of HLA-DPB1 locus, and can be easily applied and extended to other gene.

关 键 词：基因分型 序列 测序 HLA-DPB1 免疫系统 

分 类 号：R392.11[医药卫生—免疫学]