RESUMO
According to official historiography, the 10-year-old Louis XVII died in the Temple of Paris on June 8, 1795. However, public rumour spread the theory that Louis XVII escaped and that his descendants would be alive today. One such putative 'Louis XVII' was Carl Wilhelm Naundorff, who died in 1845 in Delft (the Netherlands). Comparative mitochondrial DNA (mtDNA) analysis gave evidence that his remains could not be identified as those of Louis XVII. In the present study, mtDNA analysis was performed on the heart of the young boy who died in the prison of Paris in 1795. In order to obtain the strongest evidence possible, two laboratories independently analysed the heart. The results showed that the consensus mtDNA sequence of the heart was identical to that of the maternal relatives of Louis XVII.
Assuntos
DNA Mitocondrial/genética , Pessoas Famosas , Antropologia Forense , Miocárdio/metabolismo , Feminino , Humanos , Masculino , LinhagemRESUMO
Amplification of mtDNA D-loop fragments with a length of 200 bp or more from ancient and even from fairly recent biological samples, can lead to erroneous results. This was clearly illustrated in our investigation of the putative heart of Louis XVII. By selecting different sets of primers which amplified shorter fragments of mtDNA (length 109 bp-201 bp), authentic polymorphisms could be visualised which remained undetected with the more classical primers for fragment sizes > 210 bp. Here we have extended those findings to other biological materials. A competitive PCR assay for quantitation of the amount of mtDNA for different fragment lengths, using a 10 bp deletion construct, was applied to ancient material and on a set of hairs of various ages of sampling (1966 up to the present). The results showed that DNA degradation started a few years after sampling. In the DNA extracts of the older hair shafts (1983-1995), the proportion of the number of short fragments to the number of long fragments is on average 4 in contrast to the most recent hair shafts. The numbers of amplifiable mtDNA copies for the hairs from 1975 and older were too small to show a clear difference. Use of long PCR fragments in such cases can yield misleading results. Use of short PCR fragments for the analysis of mtDNA from shed hair, in combination with a competitive PCR assay to determine the state of degradation, should improve the reliability of forensic mtDNA analysis considerably.
Assuntos
DNA Mitocondrial/genética , Pessoas Famosas , Antropologia Forense , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , DNA Mitocondrial/análise , França , Cabelo/química , História do Século XVIII , Humanos , Masculino , Miocárdio/química , Oligonucleotídeos , Linhagem , Polimorfismo Genético , Reprodutibilidade dos Testes , Extratos de Tecidos/análiseRESUMO
Carl Wilhelm Naundorff was buried in 1845 in Delft as Louis Charles, Duc de Normandie, 'Louis XVII'. However, the son of Louis XVI and Marie-Antoinette-Louis XVII--officially died in the Temple of Paris in 1795. In order to resolve the identity of Naundorff, mitochondrial DNA (mtDNA) D-loop sequences of his remains were compared with the sequences obtained from the hairs of two sisters of Marie-Antoinette, Marie-Antoinette herself, and with the sequences obtained from DNA samples of two living maternal relatives. The mtDNA sequence of a bone sample from Naundorff showed two nucleotide differences from the sequences of the three sisters and four differences from the sequences of living maternal relatives. Based on this evidence it becomes very unlikely that Naundroff is the son of Marie-Antoinette.
Assuntos
DNA Mitocondrial/genética , Pessoas Famosas , Antropologia Forense , Sequência de Bases , Sequência Consenso , Feminino , França , Humanos , Masculino , Linhagem , Polimorfismo Genético , Cromossomo YRESUMO
Mitochondrial DNA (mtDNA) sequencing is a powerful and sensitive method to identify the donor of shed hairs found at a crime scene. Because of the low amounts of DNA in shed hair and the sensitivity of PCR, contaminating cells (e.g. saliva, blood), sometimes present on these hairs, will be co-amplified. This will result in ambiguous sequencing results and might even lead to erroneous exclusions of suspects. We have evaluated a strategy for effectively removing saliva and blood contamination from hair samples. Unambiguous mtDNA results were obtained by incubating the hair samples in a differential lysis buffer (which contains no DTT) prior to DNA extraction. Since the nuclear DNA of the hair root is affected, this procedure should be restricted to hair shaft proportions.
