Danielson Phillip B, Sun Hong-Yu, Melton Terry, Kristinsson Richard
University of Denver, Department of Biological Sciences, 2190 E. Iliff Avenue Rm 102, Denver, CO 80208, USA.
Forensic Sci Int Genet. 2007 Jun;1(2):148-53. doi: 10.1016/j.fsigen.2007.02.008. Epub 2007 Mar 12.
Mitochondrial DNA (mtDNA) sequencing can provide crucial information to forensic investigators when the quantity and quality of DNA would otherwise be limiting. The difficulty of analyzing mtDNA mixtures, however, has been a significant obstacle to its broader use in forensics. Denaturing high-performance liquid chromatography (DHPLC) in combination with direct sequencing makes it possible to determine the linkage phase of individual amplicons in a mixture. The reliability of the approach is based, in part, on the strong correlation between a change in the relative quantities of different DNA amplicons in a given mixture versus a change in the relative electrophoretic peak heights at mixed base positions. Using standard operating procedures previously validated for use in forensic laboratories, this approach enables sequence-specific fractionation of natural (heteroplasmic) or situational (multi-contributor) DNA mixtures prior to direct sequencing. As a novel approach for the rapid and accurate analysis of DNA mixtures, DHPLC may aid criminal investigators by making it possible to obtain definitive mitochondrial DNA results from otherwise challenging samples.
当DNA的数量和质量受到限制时，线粒体DNA（mtDNA）测序可为法医调查人员提供关键信息。然而，分析mtDNA混合物的困难一直是其在法医学中更广泛应用的重大障碍。变性高效液相色谱（DHPLC）与直接测序相结合，使得确定混合物中单个扩增子的连锁相成为可能。该方法的可靠性部分基于给定混合物中不同DNA扩增子相对数量的变化与混合碱基位置处相对电泳峰高的变化之间的强相关性。使用先前已在法医实验室验证的标准操作程序，这种方法能够在直接测序之前对天然（异质性）或情境性（多贡献者）DNA混合物进行序列特异性分离。作为一种快速准确分析DNA混合物的新方法，DHPLC可以通过从原本具有挑战性的样本中获得明确的线粒体DNA结果来帮助刑事调查人员。
