Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health Sciences, University of Copenhagen, Frederik V's Vej 11, DK-2100 Copenhagen, Denmark.
Forensic Sci Int Genet. 2012 Mar;6(2):e72-4. doi: 10.1016/j.fsigen.2011.08.002. Epub 2011 Sep 7.
In this study, we tested the first commercial kit with insertion/deletion (indel) polymorphisms, the Mentype(®) DIPplex PCR Amplification Kit (DIPplex kit). A total of 30 biallelic autosomal indels and Amelogenin were amplified with the DIPplex kit. All loci were amplified in one PCR multiplex and all amplicon lengths were shorter than 160 bp. Full indel profiles were generated from as little as 100 pg of DNA. A total of 117 individuals from Danish paternity cases were successfully typed. No deviation from Hardy-Weinberg equilibrium was observed for any of the indels. The combined mean match probability was 3.3 × 10(-13), the mean paternity exclusion probability was 99.7% and the typical paternity indices for trios and duos were 2350 and 165, respectively. Furthermore, we typed five highly degraded DNA samples with the DIPplex kit, the AmpFlSTR(®) SGM Plus kit and the AmpFlSTR(®) SEfiler Plus kit. Full indel profiles were obtained with the DIPplex kit, whereas only partial profiles were obtained with the STR kits. In general, the DIPplex kit performed well and it would be a valuable assay for forensic genetic testing, especially in crime cases with partially degraded DNA or low amounts of template DNA. However, some difficulties with pull-ups were observed at DNA concentrations of 1000 pg. Rearrangement of the allele windows by changing the lengths of some of the PCR primers would greatly improve the assay, and more robustness towards higher amounts of DNA would allow the use of the DIPplex kit without prior quantification of the samples.
在这项研究中，我们测试了第一个带有插入/缺失（indel）多态性的商业试剂盒，即 Mentype(®) DIPplex PCR 扩增试剂盒（DIPplex 试剂盒）。该 DIPplex 试剂盒共扩增了 30 个双等位常染色体 indel 和 Amelogenin。所有位点均在一个 PCR 多重扩增中扩增，所有扩增子长度均短于 160 bp。仅从 100 pg DNA 即可生成完整的 indel 图谱。成功对来自丹麦亲子鉴定案例的 117 个人进行了分型。未观察到任何 indel 偏离 Hardy-Weinberg 平衡。合并的平均匹配概率为 3.3×10(-13)，平均亲权排除概率为 99.7%，典型的三联体和二联体亲权指数分别为 2350 和 165。此外，我们使用 DIPplex 试剂盒、AmpFlSTR(®) SGM Plus 试剂盒和 AmpFlSTR(®) SEfiler Plus 试剂盒对五个高度降解的 DNA 样本进行了分型。DIPplex 试剂盒获得了完整的 indel 图谱，而 STR 试剂盒仅获得了部分图谱。总体而言，DIPplex 试剂盒性能良好，将是法医遗传学检测的宝贵检测方法，尤其是在 DNA 部分降解或模板 DNA 量低的犯罪案件中。然而，在 DNA 浓度为 1000 pg 时，观察到一些与 Pull-ups 相关的困难。通过改变一些 PCR 引物的长度来重新排列等位基因窗口，将极大地改进该检测方法，并且提高对更高量 DNA 的鲁棒性，将允许在不事先对样本进行定量的情况下使用 DIPplex 试剂盒。
