Parson Walther, Brandstätter Anita, Alonso Antonio, Brandt Nathalie, Brinkmann Bernd, Carracedo Angel, Corach Daniel, Froment Olivier, Furac Ivana, Grzybowski Tomasz, Hedberg Karin, Keyser-Tracqui Christine, Kupiec Tomasz, Lutz-Bonengel Sabine, Mevag Bente, Ploski Rafal, Schmitter Hermann, Schneider Peter, Syndercombe-Court Denise, Sørensen Eric, Thew Heather, Tully Gillian, Scheithauer Richard
Institute of Legal Medicine, Medical University of Innsbruck, Muellerstrasse 44, Innsbruck 6020, Austria.
Forensic Sci Int. 2004 Jan 28;139(2-3):215-26. doi: 10.1016/j.forsciint.2003.11.008.
This paper presents an overview of the organisation and the results of the collaborative exercises (CE) of the European DNA Profiling (EDNAP) Group's mitochondrial DNA population database project (EMPOP). The aim of the collaborative exercises was to determine whether uniformity of mtDNA sequencing results could be achieved among different laboratories. These were asked to sequence either the complete mtDNA control region or the two hypervariable regions HVI (16024-16365) and HVII (73-340) from DNA extracts, buccal swabs or bloodstains, proceeding in accordance with the protocol and strategies used in each individual laboratory. The results of the collaborative exercises were employed to identify possible sources of errors that could arise during the analysis and interpretation of mtDNA profiles. These findings were taken as a basis to tentatively make suitable arrangements for the construction of a high quality mtDNA database. One hundred fifty mtDNA profiles were submitted to the evaluating laboratory, and disaccording profiles were classified into four groups corresponding to the source of error: clerical errors, sample mix-ups, contaminations and discrepancies with respect to the mtDNA nomenclature. Overall, 14 disaccording haplotypes (16 individual errors) were observed. The errors included 10 clerical errors, 3 interpretation problems, 2 cases of sample mix-up and 1 case of point heteroplasmic mixture, where the 2 sequencing reactions brought inconsistent base calls. This corresponds to an error rate of 10.7% in a virtual mtDNA database consisting of the collaborative exercise results. However, this estimate is still conservative compared to conclusions drawn by authors of meanwhile numerous publications critically reviewing published mtDNA population databases. Our results and earlier published concerns strongly emphasize the need for appropriate safety regulations when mtDNA profiles are compiled for database purposes in order to accomplish the high standard required for mtDNA databases that are used in the forensic context.
本文概述了欧洲DNA分型（EDNAP）集团线粒体DNA群体数据库项目（EMPOP）的协作实验（CE）的组织情况及结果。协作实验的目的是确定不同实验室之间是否能实现线粒体DNA测序结果的一致性。要求各实验室按照各自使用的方案和策略，对DNA提取物、口腔拭子或血迹中的完整线粒体DNA控制区或两个高变区HVI（16024 - 16365）和HVII（73 - 340）进行测序。协作实验的结果被用于识别线粒体DNA图谱分析和解读过程中可能出现的错误来源。这些发现被作为初步为构建高质量线粒体DNA数据库做出适当安排的依据。150个线粒体DNA图谱被提交至评估实验室，不一致的图谱被分为与错误来源相对应的四组：文书错误、样本混淆、污染以及线粒体DNA命名法方面的差异。总体而言，观察到14个不一致的单倍型（16个个体错误）。这些错误包括10个文书错误、3个解读问题、2例样本混淆以及1例点异质性混合，其中2次测序反应得出了不一致的碱基调用结果。在由协作实验结果构成的虚拟线粒体DNA数据库中，这对应的错误率为10.7%。然而，与众多批判性审视已发表的线粒体DNA群体数据库的作者得出的结论相比，这一估计仍然较为保守。我们的结果以及早期发表的相关担忧强烈强调，为数据库目的编制线粒体DNA图谱时，需要有适当的安全规定，以达到法医背景下使用的线粒体DNA数据库所需的高标准。
