Internal validation and improvement of mitochondrial genome sequencing using the Precision ID mtDNA Whole Genome Panel
| Autor: | Martina Rigato, Giovanni Vazza, Christian Faccinetto, Gianluca Casamassima, Donata Luiselli, Cecilia Salvoro, Gianluca Margiotta, Stefania Sarno, Alberto Marino, Nicola Staiti, Sara De Fanti, Daniele Sabbatini, Patrizia Serventi |
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| Přispěvatelé: | Faccinetto C., Sabbatini D., Serventi P., Rigato M., Salvoro C., Casamassima G., Margiotta G., De Fanti S., Sarno S., Staiti N., Luiselli D., Marino A., Vazza G. |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Mitochondrial DNA
Whole mitochondrial genome Forensic biology Method Paper Computational biology Forensic genetics Biology Ion Torrent DNA Mitochondrial Genome Pathology and Forensic Medicine symbols.namesake Humans Internal validation Sanger sequencing Massive parallel sequencing High-Throughput Nucleotide Sequencing Reproducibility of Results Sequence Analysis DNA Forensic genetic Ion semiconductor sequencing Heteroplasmy Haplotypes NGS Genome Mitochondrial symbols |
| Zdroj: | International Journal of Legal Medicine |
| Popis: | With the recent advances in next-generation sequencing (NGS), mitochondrial whole-genome sequencing has begun to be applied to the field of the forensic biology as an alternative to the traditional Sanger-type sequencing (STS). However, experimental workflows, commercial solutions, and output data analysis must be strictly validated before being implemented into the forensic laboratory. In this study, we performed an internal validation for an NGS-based typing of the entire mitochondrial genome using the Precision ID mtDNA Whole Genome Panel (Thermo Fisher Scientific) on the Ion S5 sequencer (Thermo Fisher Scientific). Concordance, repeatability, reproducibility, sensitivity, and heteroplasmy detection analyses were assessed using the 2800 M and 9947A standard control DNA as well as typical casework specimens, and results were compared with conventional Sanger sequencing and another NGS sequencer in a different laboratory. We discuss the strengths and limitations of this approach, highlighting some issues regarding noise thresholds and heteroplasmy detection, and suggesting solutions to mitigate these effects and improve overall data interpretation. Results confirmed that the Precision ID Whole mtDNA Genome Panel is highly reproducible and sensitive, yielding useful full mitochondrial DNA sequences also from challenging DNA specimens, thus providing further support for its use in forensic practice. Supplementary Information The online version contains supplementary material available at 10.1007/s00414-021-02686-w. |
| Databáze: | OpenAIRE |
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