运用焦磷酸测序技术进行法医线粒体编码区分析以提高鉴别能力。

Forensic mitochondrial coding region analysis for increased discrimination using pyrosequencing technology.

作者信息

Andréasson Hanna, Nilsson Martina, Styrman Hanna, Pettersson Ulf, Allen Marie

机构信息

Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.

出版信息

Forensic Sci Int Genet. 2007 Mar;1(1):35-43. doi: 10.1016/j.fsigen.2006.10.002. Epub 2006 Nov 29.

DOI:10.1016/j.fsigen.2006.10.002

PMID:19083726

Abstract

Analysis of mitochondrial DNA (mtDNA) is very useful when nuclear DNA analysis fails due to degradation or insufficient amounts of DNA in forensic analysis. However, mtDNA analysis has a lower discrimination power compared to what can be obtained by nuclear DNA (nDNA) analysis, potentially resulting in multiple individuals showing identical mtDNA types in the HVI/HVII region. In this study, the increase in discrimination by analysis of mitochondrial coding regions has been evaluated for identical or similar HVI/HVII sequences. A pyrosequencing-based system for coding region analysis, comprising 17 pyrosequencing reactions performed on 15 PCR fragments, was utilised. This assay was evaluated in 135 samples, resulting in an average read length of 81 nucleotides in the pyrosequencing analysis. In the sample set, a total of 52 coding region SNPs were identified, of which 18 were singletons. In a group of 60 samples with 0 or 1 control region difference from the revised Cambridge reference sequence (rCRS), only 12 samples could not be resolved by at least two differences using the pyrosequencing assay. Thus, the use of this pyrosequencing-based coding region assay has the potential to substantially increase the discriminatory power of mtDNA analysis.

摘要

在法医分析中，当由于DNA降解或DNA量不足导致核DNA分析失败时，线粒体DNA（mtDNA）分析非常有用。然而，与核DNA（nDNA）分析相比，mtDNA分析的鉴别能力较低，这可能导致多个个体在HVI/HVII区域表现出相同的mtDNA类型。在本研究中，针对相同或相似的HVI/HVII序列，评估了通过分析线粒体编码区提高鉴别能力的情况。使用了一种基于焦磷酸测序的编码区分析系统，该系统包括对15个PCR片段进行17次焦磷酸测序反应。对135个样本进行了该检测，焦磷酸测序分析的平均读长为81个核苷酸。在样本集中，共鉴定出52个编码区单核苷酸多态性（SNP），其中18个是单例。在一组与修订的剑桥参考序列（rCRS）在控制区有0或1个差异的60个样本中，使用焦磷酸测序检测，只有12个样本不能通过至少两个差异来区分。因此，使用这种基于焦磷酸测序的编码区检测方法有可能大幅提高mtDNA分析的鉴别能力。