Analysis of minisatellite mutation for performing a highly reliable paternity test without false exclusion

分析小卫星突变，以进行高度可靠的亲子鉴定，而不会出现错误排除

• 批准号: 12670399
• 负责人: TAMAKI Keiji
• 金额: $ 2.18万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670399/
• 关键词: minisatellite; MVR-PCR; minisatellite mutation; forensic application; paternity test

Minisatellite variant repeat (MVR) mapping using the polymerase chain reaction (PCR) was devised to map the interspersion pattern of subtle variant repeats along minisatellite tandem arrays. MVR-PCR has revealed enormous diversity of allele structures at, several loci, far more than can be resolved by allele length analysis. We showed that the application of MVR-PCR resulted in higher paternity probabilities than for a set of 12 other classic DNA markers. Hypervariable minisatellites like MS32 and MS31A can however show significant germline mutation rates to new length alleles which can generate false exclusions in paternity cases although paternity cases showing mutant paternal alleles at more than one locus will be rare when several MVR loci are examined. Detailed knowledge of mutation processes coupled with MVR analysis of allele structure can help distinguish mutation from non-paternity. We show how similar mutant alleles are to their progenitors using both real and simulated data, … More and demonstrate how MVR-PCR can be used to identify mutant paternal allele in paternity cases showing apparent exclusions.We analyse minisatellite B6.7 locus in the Japanese population. Allele size distributions showed that Japanese retain extensive allele length variability but have significantly smaller alleles compared to north Europeans. Ninety-two Japanese alleles were further characterised by MVR-PCR. These alleles showed a wide variety of internal MVR structures with most alleles observed only once in the sample. The true heterozygosity is estimated at 99.95%, with well in excess of 2000 different alleles existing in the Japanese population. Dot matrix analysis showed that groups of related alleles sharing structural motifs could be identified within Japanese and in north Europeans, and that these groups are population-specific with no examples of significant similarity between any Japanese and north European alleles. Minisatellite B6.7 therefore shows huge allele variability and fast repeat turnover in Japanese as well as north European populations, and provides novel lineage markers for exploring very recent events in human population history. Less

设计了使用聚合酶链反应（PCR）的微型变体重复（MVR）映射，以绘制沿着微型变体重复序列的散布模式，沿着微型卫星串联阵列。 MVR-PCR揭示了等位基因结构的巨大多样性，几个局部，远远超过了通过等位基因长度分析可以解决的。我们表明，与其他12个其他经典DNA标记相比，MVR-PCR的应用导致了更高的亲子关系可能性。但是，像MS32和MS31A这样的高变量迷你卫星可以显示出对新长度等位基因的生殖线突变速率，尽管在检查了几个MVR基因座时很少有一个以上的基因座，但在亲子鉴定病例中可能会产生错误的排除。对突变过程以及等位基因结构的MVR分析的详细知识可以帮助区分突变与非及格性。我们展示了使用真实数据和模拟数据的相似突变等位基因与它们的祖细胞的相似性，并证明了如何使用MVR-PCR来鉴定出明显排除的亲子关系案例中的突变亲属等级。等位基因的大小分布表明，日本人保留了广泛的等位基因长度可变性，但与北欧人相比，等位基因明显小。 MVR-PCR进一步表征了92个日本等位基因。这些等位基因显示出多种内部MVR结构，大多数等位基因在样品中仅观察到一次。真正的杂合性估计为99.95％，在日本人口中存在超过2000种不同的等位基因。 DOT矩阵分析表明，可以在日本和北欧人中确定相关等位基因共享结构图案的组，并且这些群体是特定于人群的，没有任何日本和北欧等位基因之间的显着相似之处。因此，Minisatellite B6.7在日本和北欧人群中显示出巨大的等位基因变异性和快速的重复失误，并为探索人口历史上最近事件提供了新颖的血统标记。较少的

项目成果

T.Yamamoto: "The application of minisatellite variant repeat mapping by PCR (MVR-PCR) in a paternity case showing false exclusion due to STR mutation"J. Forensic Sci.. 46. 374-378 (2001)

T.Yamamoto：“通过 PCR (MVR-PCR) 进行小卫星变异重复定位在亲子鉴定案例中的应用，显示由于 STR 突变导致的错误排除”J.

K.Tamaki: "Distinguishing minisatellite mutation from non-paternity by MVR-PCR"Forensic Sci.Int.. 113. 56-62 (2000)

K.Tamaki：“通过 MVR-PCR 区分小卫星突变和非亲子突变”Forensic Sc​​i.Int.. 113. 56-62 (2000)

K.Tamaki: "Distinguishing minisatellite mutation from non-paternity by MVR-PCR"Forensic Sci. Int.. 113. 55-62 (2000)

K.Tamaki：“通过 MVR-PCR 区分小卫星突变和非亲子突变”《法医科学》。

M.Ito et al.: "Rapid detection of minisatellite alleles by allele-specific MVR-PCR using fluorescent labeled primers (in Japanese)"DNA takei. 9. 268-271 (2001)

M.Ito 等人：“使用荧光标记引物通过等位基因特异性 MVR-PCR 快速检测小卫星等位基因（日语）”DNA takei。

T.Mizukoshi et al.: "Allelic structures at hypervariable minisatellite B6.7 in Japanese show population specificity"J.Hum.Genet. 47(6). 232-238 (2002)

T.Mizukoshi 等人：“日本高变小卫星 B6.7 的等位基因结构显示群体特异性”J.Hum.Genet。