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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.

本研究试图建立一种适合于法医学个人识别和亲子鉴定的单核苷酸多态性（SNPs）基因型检测方法。首先，我们测试了几种方法来搜索哪一种是最好的。等位基因特异性PCR法便于基因座筛选，SnapShot法适合于多重体系。然后，我们选择了高度多态性的51个SNP位点的亚洲人口以外的区域的基因从那些在SNP联盟。其中，39个位点与32名日本健康人的x^2检验结果符合Hardy-Weinberg平衡。这39个基因座在日本人中具有高度多态性，其中22个基因座的频率低于0.4，14个基因座的频率高于0.3。我们现在正在建立一个新的多重SNP分型系统，这12个位点分布在不同的染色体上，使用相同的技术，我们已经成功地在新设计的STR多重系统。我们还试图建立一种高灵敏度的方法，用于DNA高度降解的微小和古老的标本。在STR分型中，由于扩增不足，常得阴性结果。大多数STR基因座的PCR产物为150-300个碱基，而SNP基因座的PCR产物可缩短至100个碱基以下。因此，SNP位点在降解DNA的高效扩增方面具有很大的优势。几个SNP位点的高灵敏度检测方法已基本完成。