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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（single nucleotide polymorphisms，SNP）是人类基因组中数量最多的DNA标记，已引起许多研究领域的关注，本研究尝试建立一种基于多重单核苷酸引物延伸反应和毛细管电泳的SNP分型方法。利用该方法可建立ABO血型基因分型方法、HLA-DRB 1基因分型方法、Y染色体多重SNPs分型方法以及实现每条染色体37重SNPs分型的高效方法。主要研究结果如下：1.用该方法测定了103例日本人的ABO基因型，其等位基因频率与文献报道的结果一致。我们将此方法应用于个案样本，并成功地确定了骨骼，牙齿，肌肉，器官，指甲和精液污染的阴道内容物的ABO基因型.我们检测了159名日本男性Y染色体上15个不同的SNP位点，检测到13种单倍型。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。该方法可应用于个人身份识别和专利性检验.本方法可用于HLA-DRB 1基因分型，特别适用于混合型人群的分型.我们建立了一种有效的方法，实现了对每条染色体上的SNPs进行37重分型。该方法有望成为分析法医标本的有效手段，并为我们提供了高的分辨力。