一种以测序为基础的基因分型方法的建立

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个随机挑选的中国南方人的HLA－DPB1基因进行了基因分型，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。

A Sequence-based Typing Method for HLA-DPB1 Genotyping

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.