权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Construction and evaluation for the multiplex typing system useful for highly degraded mixed stain specimens

适用于高度降解混合染色样本的多重分型系统的构建和评估

基本信息

批准号：
15590577
负责人：
YAMAMOTO Toshimichi
金额：
$ 2.24万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590577/
关键词：
STR Mixed stain Multiplex Degraded DNA Japanese Genetic polymorphism

项目摘要

A newly commercially released kit "Penta BEC Multiplex kit" was evaluated for its usefulness in a Japanese population. The allele frequencies at those loci were calculated from 300 unrelated Japanese. No significant deviations from Hardy-Weiberg equilibrium(HWE) were observed using three tests at the three loci. The power of discrimination(PD) values for all the three loci were more than 0.90,and the accumulated PD and PE(power of exclusion) were 0.99991 and 0.96531,respectively in this multiplex system. The stutter peak percentages at the three loci were also calculated, those of these pentanucleotide repeat STR were lower than those of tetrranucleotide repeat used generally included in a commercially released kit. Furthermore, the stutter percentages showed the positive correlation with the repeat numbers of the alleles, almost all of them distributed in the range of the mean±3 SD at each allele. Therefore, it was suggested that t the mean±3 SD at each allele could de a standard to distinguish between the stutter peak and the minor peak originated fom a mixed stain sample.We have almost completed to construct a novel quadruplex for pentanucleotide repeat STR loci (Penta, E, B and D, and D10S2325), in which the primer sets were newly designed, but a few of the allelic ladder markers was not finished to confirm the sequences. As a part of the evaluation of the system, the stutter peak percentages were calculated. The allele frequencies at those loci were calculated from three regional Japanese populations (Akita, Nagoya and Oita). No.significant deviations from HWE were observed using three tests at the all loci. The observed heterozygosity and PD values for all the three loci in the three populations were more than 0.77 and 0.92,respectively. Compared pairwisely among the allele frequency distributions of the three loci, two loci between Akita and Oita, one locus between Akita and Nagoya, and between Nagoya-Oita were significantly different (P<0.05).

评估了新商业发布的试剂盒“Penta BEC Multiplex试剂盒”在日本人群中的有效性。在这些位点的等位基因频率计算从300无关的日本人。在三个位点使用三个测试没有观察到显著偏离Hardy-Weiberg平衡（HWE）。在该复合体系中，3个基因座的识别能力（PD）均大于0.90，累积PD和PE分别为0.99991和0.96531。同时计算了三个位点的突变峰百分比，这些五核苷酸重复序列的突变峰百分比低于市售试剂盒中通常使用的四核苷酸重复序列的突变峰百分比。口吃率与等位基因重复数呈正相关，几乎都分布在每个等位基因的平均值± 3SD范围内。因此，每个等位基因的平均值± 3SD可以作为区分混合染色样品中的断续峰和次要峰的标准。我们基本完成了五核苷酸重复序列STR基因座的四链体构建（Penta、E、B和D以及D 10 S2325），其中引物组是新设计的，但一些等位基因阶梯标记尚未完成以确认序列。作为系统评价的一部分，计算了断续峰百分比。这些基因座的等位基因频率计算从三个地区的日本人口（秋田，名古屋和大分）。在所有位点使用三次试验未观察到与HWE的显著偏离。3个群体的3个基因座的杂合度和PD值分别大于0.77和0.92。两两比较，秋田与大分、秋田与名古屋、名古屋与大分之间的等位基因频率分布差异显著（P<0.05）。