Morling N, Hansen H E
Institute of Forensic Genetics, University of Copenhagen, Denmark.
Int J Legal Med. 1993;105(4):189-96. doi: 10.1007/BF01642792.
Paternity testing using DNA polymorphism of variable numbers of tandem repeat (VNTR) regions with restriction fragment length polymorphism (RFLP) was implemented. HinfI-digested DNA was separated by electrophoresis in agarose gels and hybridized with radiolabelled probes detecting the VNTR-systems D2S44 (YNH24), D5S43 (MS8), D7S21 (MS31), D7S22 (g3), and D12S11 (MS43a). The intra gel variability of 970 duplicate investigations on the same gel of DNA from 122 individuals showed no differences exceeding 1.25 mm between the positions of the corresponding DNA fragments. The comparison of 1,624 DNA fragments from 342 mother/child pairs showed only one difference above 1.25 mm which was interpreted as a mutation. Based on these observations, we decided to consider an intra gel difference above 1.25 mm between the non-maternal DNA fragment of the child and the nearest DNA fragment of the putative father as an exclusion in paternity testing. This matching criterion was used for the comparisons of 1,197 DNA fragment differences in 247 pairs of children and putative fathers who had not been excluded by conventional marker systems. In all of these cases, the migration differences between the DNA fragments of non-excluded men and the DNA fragments of the children were less than 1.25 mm except in 6 cases (0.5%). The man/child differences in all of 227 false trios exceeded 1.25 mm in 2 or more of the 5 VNTR systems investigated. Matching criteria for inter gel comparisons in paternity testing were established. The frequency distribution of HinfI digested DNA fragments of the 5 VNTR systems in 650 unrelated Danes is presented and the raw data is available.
采用限制性片段长度多态性(RFLP)技术,利用可变数目串联重复序列(VNTR)区域的DNA多态性进行亲子鉴定。用HinfI酶消化的DNA在琼脂糖凝胶中进行电泳分离,然后与检测VNTR系统D2S44(YNH24)、D5S43(MS8)、D7S21(MS31)、D7S22(g3)和D12S11(MS43a)的放射性标记探针杂交。对来自122个人的DNA在同一凝胶上进行的970次重复检测的凝胶内变异性显示,相应DNA片段位置之间的差异不超过1.25毫米。对342对母子对的1624个DNA片段进行比较,仅发现一处差异超过1.25毫米,被解释为突变。基于这些观察结果,我们决定在亲子鉴定中将孩子的非母系DNA片段与推定父亲最近的DNA片段之间凝胶内差异超过1.25毫米视为排除亲子关系。该匹配标准用于比较247对未被传统标记系统排除的孩子与推定父亲之间的1197个DNA片段差异。在所有这些案例中,除6例(0.5%)外,未被排除的男性的DNA片段与孩子的DNA片段之间的迁移差异均小于1.25毫米。在所有227个假三联体中,在研究的5个VNTR系统中的2个或更多系统中,男性/孩子差异超过1.25毫米。建立了亲子鉴定中凝胶间比较的匹配标准。给出了650名不相关丹麦人5个VNTR系统的HinfI消化DNA片段的频率分布,原始数据也可获取。