Forensic molecular biological applications using single nucleotide polymorphisms (SNPs) regions

使用单核苷酸多态性 (SNP) 区域的法医分子生物学应用

基本信息

批准号：
12557041
负责人：
KATSUMATA Yoshinao
金额：
$ 8.32万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2003
项目状态：
已结题

项目摘要

In this study, we tried to establish the appropriate method to detect the genotypes of single nucleotide polymorphisms (SNPs) which are useful for forensic practice such as personal identification and paternity testing. At first, we tested several methods to search which is the best. We found that the allele specific PCR method is convenient for locus selection, and the SnapShot method is suitable for multiplex system. Then, we selected highly polymorphic 51 SNP loci in an Asian population being outside the region of the gene from those listed in the SNP Consortium. Among them, 39 loci were found to be in the Hardy-Weinberg equilibrium by x^2 method with 32 unrelated healthy Japanese. These 39 loci are highly polymorphic in Japanese ; 22 loci show the lower frequencies over than 0.4, and 14 show them over than 0.3. We are now establishing a new multiplex SNP typing system with the 12 out of these 14 loci, which distribute on different chromosome using the same techniques by which we have succeeded in the newly devised STR multiplex system. We also tried to establish a highly sensitive method for the minute and old specimens where DNA is highly degraded. We often get negative results in STR typing with such specimens because of the insufficient amplification. The PCR products of most STR loci are 150-300 bases, while those of SNP loci can be shortened to less than 100 bases. Therefore, SNP loci have a great advantage of the efficient amplification of degraded DNA. We could almost finish the development of the highly sensitive method for several SNP loci.

在这项研究中，我们试图建立适当的方法来检测单核苷酸多态性（SNP）的基因型，这对于个人身份识别和亲子鉴定等法医实践非常有用。首先，我们测试了几种方法来寻找最好的。我们发现等位基因特异性PCR方法便于位点选择，而SnapShot方法适合多重系统。然后，我们从 SNP 联盟中列出的基因区域之外的亚洲人群中选择了高度多态性的 51 个 SNP 位点。其中，通过x^2方法发现39个位点与32个无关的健康日本人处于Hardy-Weinberg平衡。这 39 个基因座在日语中具有高度多态性； 22 个位点显示频率低于 0.4，14 个位点显示频率高于 0.3。我们现在正在建立一个新的多重 SNP 分型系统，其中 14 个基因座中的 12 个使用我们在新设计的 STR 多重系统中成功的相同技术，这些基因座分布在不同的染色体上。我们还尝试建立一种针对 DNA 高度降解的微小和陈旧样本的高灵敏度方法。由于扩增不足，我们经常在使用此类样本进行 STR 分型时得到阴性结果。大多数STR位点的PCR产物为150-300个碱基，而SNP位点的PCR产物可以缩短至100个碱基以下。因此，SNP位点对于降解DNA的高效扩增具有很大的优势。我们已经基本完成了针对多个SNP位点的高灵敏方法的开发。