Establishment of high-sensitive multiplex typing systems for Y-STRs and its application to a very small amount of specimens

Y-STR高灵敏多重分型系统的建立及其在极少量标本中的应用

• 批准号: 13670421
• 负责人: KATSUMATA Yoshinao
• 金额: $ 2.3万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670421/
• 关键词: Y-chromosome; STR; Multiplex; Genetic polymorphism; High-sensitive detection

In the preliminary step toward the present study, two novel multiplex typing systems for autosomal STR loci were constructed. These contributions were published in the journals. One of them was specially designed to have high sensitivity of the system. In order to select the loci with higher discriminating power, the haplotypes at 14 Y-STR loci were analyzed from two regional Japanese DNA samples (207 in Honshu and 87 in Okinawa) and 117 Thai samples using the 10-plex (DYS436, DYS439, DYS435, DYS19, DYS460, Y-GATA-H4, DYS391, DYS392, DYS438, DYS437) devised by NIST in the USA and the commercially released Y-PLEX6 kit (DYS393, DYS19, DYS389, DYS390, DYS391, DYS385) because of having two loci (DYS19 and DYS391) in common in both systems. Based on these studies, we completed to construct two kinds of high-sensitive multiplex systems for highly degraded DNA samples. One was for three loci (DYS392, DYS438 and DYS393) labeled with three-colored fluorescent dyes and another was for four loci (DYS19, DYS3891, DYS438 and DYS460) labeled with three-colored ones. The latter was especially useful because it made it possible to type the 4 loci from 50 pg of template DNA at usual numbers of PCR cycles (30 cycles) and from less amount of the template by increasing the number of cycles. Since the haplotype diversity was also calculated from 94 Japanese male samples, and the value was very high with 0.972, it was indicated this system could be very useful in forensic fields from the point of discriminating power as well. These results will be submitted elsewhere in near future. By this grant, our study to construct the multiplex system has been achieved to the goal.

在本研究的第一步中，构建了两个针对常染色体 STR 基因座的新型多重分型系统。这些贡献发表在期刊上。其中之一是专门设计的，具有高灵敏度的系统。为了选择具有更高区分能力的位点，使用 10 重（DYS436、DYS439、DYS435、DYS19、DYS460、Y-GATA-H4、DYS391、 美国NIST设计的DYS392、DYS438、DYS437）和商业发布的Y-PLEX6试剂盒（DYS393、DYS19、DYS389、DYS390、DYS391、DYS385），因为两个系统中有两个共同的基因座（DYS19和DYS391）。基于这些研究，我们完成了两种针对高度降解DNA样品的高灵敏度多重系统的构建。一种是用三色荧光染料标记的三个基因座（DYS392、DYS438和DYS393），另一种是用三色荧光染料标记的四个基因座（DYS19、DYS3891、DYS438和DYS460）。后者特别有用，因为它可以在通常的 PCR 循环数（30 个循环）下从 50 pg 模板 DNA 中分型 4 个基因座，并通过增加循环数从较少量的模板中分型。由于单倍型多样性也是从94个日本男性样本中计算出来的，并且该值非常高，达到0.972，这表明该系统从区分能力的角度来看在法医学领域也非常有用。这些结果将在不久的将来提交到其他地方。通过这笔资助，我们构建多重系统的研究已经达到了目标。

项目成果

Yoshimoto T: "A novel fluorescent quadruplex STR typing system and the allele frequency distributions in a Thai population."Journal Forensic Sciences. 48(1). 116-121 (2003)

Yoshimoto T：“一种新型荧光四联体 STR 分型系统和泰国人群中的等位基因频率分布。”《法医学杂志》。

R.Uchihi: "Haplotype analysis with 14 Y-STR loci using 2 multiplex amplification and typing systems in 2 regional populations in Japan"International J. Legal Medicine. 117. 34-38 (2003)

R.Uchihi：“在日本 2 个地区人群中使用 2 个多重扩增和分型系统对 14 个 Y-STR 位点进行单倍型分析”International J. Legal Medicine。

Usuda K: "Haplotype analysis at 10 Y-STR loci using a novel multiplex PCR amplification and typing system in the Japanese and Thai populations."DNA Polymorphisms. 10. 164-167 (2002)

Usuda K：“在日本和泰国人群中使用新型多重 PCR 扩增和分型系统对 10 个 Y-STR 位点进行单倍型分析。”DNA 多态性。

H.Ohtaki: "A powerful, novel, multiplex typing system for six short tandem repeat loci and the allete frequency distributions in two Japanese regional populations"Electrophoresis. 23(19). 3332-3340 (2002)

H.Ohtaki：“一种强大的、新颖的多重分型系统，用于两个日本地区人群中的六个短串联重复基因座和等位基因频率分布”电泳。

打樋利英子: "低分子化DNA試料に有用な新しい Y-STR quadruplex タイピングシステム"DNA多型. 11(印刷中). (2003)

Rieko Uchihi：“适用于低分子 DNA 样品的新型 Y-STR 四联体分型系统”DNA 多态性（2003 年）。