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

利用聚合酶链反应（PCR）技术，设计了小卫星变异重复序列（MVR）图谱，以绘制微小变异重复序列沿着小卫星串联阵列的散布模式。MVR-PCR已经揭示了几个基因座上等位基因结构的巨大多样性，远远超过了等位基因长度分析所能分辨的。我们发现，MVR-PCR的应用导致更高的亲子鉴定概率比一组其他12个经典的DNA标记。然而，高变小卫星如MS 32和MS 31 A可以显示对新长度等位基因的显著种系突变率，这可以在亲子鉴定病例中产生错误排除，尽管当检查几个MVR基因座时，在多于一个基因座处显示突变的父系等位基因的亲子鉴定病例将是罕见的。突变过程的详细知识加上等位基因结构的MVR分析可以帮助区分突变与非亲子关系。我们使用真实的和模拟的数据来显示突变等位基因与其祖先的相似程度， ...更多信息 本文分析了日本人群中B6.7小卫星位点的基因型，并对MVR-PCR在亲权鉴定中的应用进行了探讨。等位基因大小分布表明，日本人保留广泛的等位基因长度变异，但有显着较小的等位基因相比，北欧人。通过MVR-PCR进一步表征了92个日本等位基因。这些等位基因表现出多种多样的内部MVR结构，大多数等位基因在样品中仅观察到一次。真实杂合性估计为99.95%，在日本人群中存在超过2000个不同的等位基因。点阵分析表明，共享结构基序的相关等位基因组可以在日本人和北欧人中识别，并且这些组是人群特异性的，没有任何日本人和北欧等位基因之间的显着相似性的例子。因此，小卫星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更替，并为探索人类人口历史中最近的事件提供了新的谱系标记。少

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