用ESI-TOF-MS分型技术检测线粒体DNA高变区碱基组成的多态性

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.

Detection of Polymorphisms of Base Compositions of Mitochondrial DNA Hypervariable Regions

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 （HV Ⅰ ） derived from primers covering coordinates15924-16428 and base compositions of 12 amplicons in hypervariable region 2 （HV Ⅱ ） derived from primers covering coordinates 31-576 were determined utilizing PLEX-ID. The polymorphisms of mtD- NA 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 HV I and ten segments in HV Ⅱ 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 HV Ⅰ and HV Ⅱ, 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.