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.

在本研究的初步阶段，我们构建了两个新的常染色体STR基因座多重分型系统。这些文章发表在期刊上。其中一个是专门设计的，具有高灵敏度的系统。为了选择具有较高鉴别力的位点，利用美国NIST设计的10-plex （DYS436、DYS439、DYS435、DYS19、DYS460、y - gath - h4、DYS391、DYS392、DYS438、DYS437）和已上市的Y-PLEX6试剂盒（DYS393、DYS19、DYS389、DYS390、DYS391、DYS385），对日本2个地区DNA样本（Honshu地区207个、冲绳地区87个）和117个泰国样本的14个Y-STR位点进行单倍型分析。在此基础上，我们构建了两种高度降解DNA样品的高灵敏度复用系统。一份是用三色荧光染料标记的三个基因座（DYS392、DYS438和DYS393），另一份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的模板DNA中分型4个位点，并通过增加循环数从较少的模板中分型。同时对94份日本男性样本进行了单倍型多样性计算，结果高达0.972，表明该系统在司法鉴定领域具有很强的应用价值。这些结果将在不久的将来提交到其他地方。在此基础上，实现了构建多路复用系统的研究目标。

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）。