Yao Lan, Xu Zhen, Zhao Hemiao, Tu Zheng, Liu Zhifang, Li Wanshui, Hu Lan, Wan Lihua
College of Basic Medicine, Chongqing Medical University, Chongqing 400016, PR China.
Key Laboratory of Forensic Genetics, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, PR China.
Leg Med (Tokyo). 2018 May;32:27-30. doi: 10.1016/j.legalmed.2018.02.005. Epub 2018 Mar 2.
In this study, the complete mitochondrial genome (mtGenome) of six samples from three forensic cases was sequenced using the Ion Torrent Personal Genome Machine (PGM). The analyzed samples from forensic cases included bloodstains from several materials, such as gauze, Flinder's Technology Associates (FTA) cards and swabs. The age of the samples ranged from two months to twelve years. The complete mtGenomes were amplified using the tiling sequencing strategy which divided the whole mtGenome into 162 amplicons. All amplicons were successfully recovered. A phylogenetic analysis was performed to determine the accuracy of the PGM data, and which were compared to partial Sanger-based sequencing data. The average coverage of the PGM data were above 4000× in all case samples, and 99.86% concordance was observed using both sequencing methods. In conclusion, we demonstrate the ability to recover the complete mtGenome from bloodstains with relatively poor DNA quality by PGM. Moreover, the results are concordant with Sanger sequencing data. This new method has potential use in forensic practice.
在本研究中，使用Ion Torrent个人基因组测序仪（PGM）对来自三个法医案件的六个样本的完整线粒体基因组（mtGenome）进行了测序。法医案件的分析样本包括来自多种材料的血迹，如纱布、弗林德斯技术协会（FTA）卡和拭子。样本的年龄范围为两个月至十二年。使用平铺测序策略扩增完整的mtGenome，该策略将整个mtGenome分为162个扩增子。所有扩增子均成功回收。进行了系统发育分析以确定PGM数据的准确性，并将其与基于桑格测序的部分数据进行比较。所有案件样本中PGM数据的平均覆盖率均高于4000倍，两种测序方法的一致性为99.86%。总之，我们证明了通过PGM从DNA质量相对较差的血迹中恢复完整mtGenome的能力。此外，结果与桑格测序数据一致。这种新方法在法医实践中有潜在用途。
