Development of a high throughput SNP typing method using capillary electrophoresis

使用毛细管电泳开发高通量 SNP 分型方法

基本信息

批准号：
13470102
负责人：
ISHIZU Hideo
金额：
$ 8.45万
依托单位：
OKAYAMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470102/
关键词：
DND polymorphisms Single nucleotide polymorphisms (SNPs)ABO blood groups HLA DRB1 types Sex chromosome Multiplex typing method High throughput Capillary electrophoresis 一塩基多型(SNP)一塩基多型(SNPs)

项目摘要

Single nucleotide polymorphisms (SNP), the most numerous DNA markers in the human genome, have attracted the attention in many research fields.In this research project, we have attempted to construct the method for SNP typing by a multiplexed single nucleotide primer extension reaction and capillary electrophoresys. Using this method, we could establish a method for genotyping of ABO blood groups, a method for genotyping of HLA-DRB1, a method for multiplex SNPs typing of Y chromosome, and an efficient method which realizes 37-plex typing of SNPs on each chromosome. The results obtained were as follows.1. ABO genotypes of 103 Japanese could be determined by this method, and the estimated allele frequencies were agreed with the previous reports. We applied this method to casework samples, and successfully determined ABO genotypes of bones, teeth, muscles, organs, nails, and semen-contaminated vaginal contents.2. We examined 15 different SNP loci on Y chromosome in 159 Japanese males, and detected 13 haplotypes. Gene diversity for each locus ranged from 0.025 to 0.486 and the haplotype diversity was estimated to be 0.838. This method could be applied for personal identification and patenity testing.3. We could establish a method for genotyping of HLA-DRB1 by the present method, and the method was useful particularly in the analysis of mixed specimens.4. We have establish an efficient method which realizes 37-plex typing of SNPs on each chromosome. This method holds promise as an effective means for analyzing forensic specimens and provides us with high discrimination power.

单核苷酸多态性（SNP）作为人类基因组中数量最多的DNA标记，已经引起了许多研究领域的关注。在本研究项目中，我们尝试构建通过多重单核苷酸引物延伸反应进行SNP分型的方法，毛细管电泳。利用该方法，我们可以建立ABO血型基因分型方法、HLA-DRB1基因分型方法、Y染色体多重SNP分型方法以及实现每条染色体上37重SNP分型的高效方法。。得到的结果如下： 1．该方法可测定103名日本人的ABO基因型，估计的等位基因频率与之前的报道一致。我们将此方法应用于个案样本，成功测定了骨骼、牙齿、肌肉、器官、指甲和精液污染的阴道内容物的ABO基因型。 2．我们检查了 159 名日本男性 Y 染色体上的 15 个不同的 SNP 位点，并检测到了 13 个单倍型。每个位点的基因多样性范围为 0.025 至 0.486，单倍型多样性估计为 0.838。该方法可用于个人身份识别和通畅性检测。 3．利用本方法建立了HLA-DRB1基因分型方法，特别适用于混合标本的分析。 4．我们建立了一种有效的方法，可以实现每条染色体上 SNP 的 37 重分型。该方法有望成为分析法医标本的有效手段，并为我们提供高辨别力。