Validation of the barcoding gene COI for use in forensic genetic species identification
Validation of the barcoding gene COI for use in forensic genetic species identification
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DOI：
10.1016/j.forsciint.2006.09.013
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发表时间：
2007-11-15
影响因子：
2.2
通讯作者：
Thorpe, Roger S.
中科院分区：
文献类型：
--
作者：
Dawnay, Nick;Ogden, Rob;Thorpe, Roger S.
法医学在野生动物犯罪调查中的应用通常涉及基于 DNA 序列相似性的遗传物种识别。这项工作可能会因缺乏经过验证的参考 DNA 序列数据而受到阻碍，从而导致证据与参考样本之间的匹配度较弱。 DNA 条形码的引入凸显了 mtDNA 基因、细胞色素 c 氧化酶 I (COI) 作为物种鉴定遗传标记的广泛使用。在这里，我们按照已发布的人体验证指南评估 COI 基因用于法医分析的情况。验证实验研究了再现性、异质性、混合 DNA、DNA 模板浓度、化学处理、底物变化、环境条件和热循环参数。使用 GenBank BLASTn 和 BOLD 搜索引擎进行的序列相似性搜索表明，COI 基因一致地识别存在经过验证的参考序列数据的物种。如果发生错误识别，其原因可归因于已发表数据中的错误参考序列或缺乏引物特异性。尽管在某些样品处理下观察到扩增失败，但没有证据表明所生成的序列中存在环境诱导的序列突变。模拟案例研究比较了 COT 和细胞色素 b mtDNA 基因的性能。讨论了与 COI 基因在法医物种鉴定中的实用性相关的研究结果。 (C) 2007 Elsevier Ireland Ltd. 保留所有权利。
The application of forensics to wildlife crime investigation routinely involves genetic species identification based on DNA sequence similarity. This work can be hindered by a lack of authenticated reference DNA sequence data resulting in weak matches between evidence and reference samples. The introduction of DNA barcoding has highlighted the expanding use of the mtDNA gene, cytochrome c oxidase I (COI), as a genetic marker for species identification. Here, we assess the COI gene for use in forensic analysis following published human validation guidelines. Validation experiments investigated reproducibility, heteroplasmy, mixed DNA, DNA template concentration, chemical treatments, substrate variation, environmental conditions and thermocycling parameters. Sequence similarity searches using both GenBank BLASTn and BOLD search engines indicated that the COI gene consistently identifies species where authenticated reference sequence data exists. Where misidentification occurred the cause was attributable to either erroneous reference sequences from published data, or lack of primer specificity. Although amplification failure was observed under certain sample treatments, there was no evidence of environmentally induced sequence mutation in those sequences that were generated. A simulated case study compared the performance of COT and cytochrome b mtDNA genes. Findings are discussed in relation to the utility of the COI gene in forensic species identification. (C) 2007 Elsevier Ireland Ltd. All rights reserved.