Abstract:

Objective To detect hypervariable regions of D-loop of mitochondrial DNA （mtDNA） by PCR/ESI-TOF-MS method and analyze the polymorphisms of base compositions. Method Base compositions of 12 amplicons in hypervariable region 1 （HVⅠ） derived from primers covering coordinates15924-16428 and base compositions of 12 amplicons in hypervariable region 2 （HVⅡ） derived from primers covering coordinates 31-576 were determined utilizing PLEX-ID. The polymorphisms of mtDNA among Han population in eastern China were investigated and the typing method was applied to a special duo maternity testing case. Results Base compositions of eight segments in HVⅠand ten segments in HVⅡ were found to be polymorphic. After ESI-TOF-MS analysis, it is found that mtDNA markers were important supplements to autosomal STR loci. Based on the different base compositions of HVⅠand HVⅡ, the questioned mother was excluded as the biological mother. Conclusion The mass spectrometry-based method for mitochondrial profiling have promising prospects. It can provide technical support for drawing correct conclusions in some special cases.

Key words: forensic genetics, mtDNA, ESI-TOF-MS, PLEX-ID, SNP

摘要：

目的 用ESI-TOF-MS分型技术检测线粒体DNA的D环高变区，通过碱基组成分析其多态性。方法 在PLEX-ID技术平台上，分别对mtDNA 高变区1 （HVⅠ，15924-16428nt）和 mtDNA 高变区Ⅱ（HVⅡ， 31-576 nt）进行碱基组成分析，考察mtDNA在华东汉族人群的多态性，并将该技术应用于一例特殊的亲子鉴定案件。结果 用ESI-TOF-MS分型技术检测线粒体DNA，在高变区Ⅰ的8个区段检见碱基组成的多态性，在mtDNA 高变区Ⅱ的10个区段检见多态性。在所应用的亲子鉴定案例中，线粒体DNA标记成了常染色体STR基因座的重要补充，经高变区Ⅰ和高变区Ⅱ的碱基组成检测，最后排除了非母。结论  ESI-TOF-MS检测mtDNA的技术具有良好的应用前景，在一些特殊的案件中，该法可为最终获得可靠鉴定结论提供技术支撑。

关键词: 法医遗传学, 线粒体DNA, ESI-TOF-MS, PLEX-ID, SNP