Construction and evaluation for the multiplex typing system useful for highly degraded mixed stain specimens
| Project/Area Number |
15590577
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | Nagoya University |
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Principal Investigator |
YAMAMOTO Toshimichi Nagoya University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (50260592)
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| Co-Investigator(Kenkyū-buntansha) |
KATSUMATA Yoshinao Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (30109326)
UCHIHI Rieko Nagoya University, Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (20223571)
TAMAKI Keiji Kyoto University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (90217175)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | STR / Mixed stain / Multiplex / Degraded DNA / Japanese / Genetic polymorphism |
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| Research Abstract |
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).
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Report
(3 results)
Research Products
(22 results)
