Abstract
Analysis of mtDNA variation using Sanger sequencing does not allow accurate quantification of the components of mtDNA mixtures. An alternative method to determine the specific mixture ratios in samples displaying heteroplasmy, consisting of DNA contributions from several individuals, or containing contamination would therefore be valuable. A novel quantification system for mtDNA mixture analysis has been developed based on pyrosequencing technology, in which the linear relationship between incorporated nucleotides and released light allows quantification of the components of a sample. Within five polymerase chain reaction fragments, seven variable positions in the mtDNA control and coding region were evaluated using this quantification analysis. For all single nucleotide polymorphisms quantified in this study, a linear relationship was observed between the measured and expected mixture ratios. This mtDNA quantification assay is an easy to use, fast and accurate quantification system, with the ability to resolve and interpret major and minor mtDNA components in forensic mixture samples.
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Acknowledgements
This project was supported by grants from Kjell and Märta Beijers Foundation, the Göran Gustafsson Foundation, the Swedish Research Council (Medicine) and VINNOVA (Swedish Agency for Innovation Systems).
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H. Andréasson and M. Nilsson have contributed equally to the work.
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Andréasson, H., Nilsson, M., Budowle, B. et al. Quantification of mtDNA mixtures in forensic evidence material using pyrosequencing. Int J Legal Med 120, 383–390 (2006). https://doi.org/10.1007/s00414-005-0072-8
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DOI: https://doi.org/10.1007/s00414-005-0072-8