- •DNA analysis from skeletal remains is crucial for missing person identification.
- •STR profiling is only suitable if ante-mortem samples or relatives are available.
- •If unavailable, appearance DNA prediction can provide leads to find relatives.
- •The HIrisPlex system allows eye and hair colour prediction from DNA.
- •We find HIrisPlex suitable for analysing remains of World War II victims.
Retrieving information about externally visible characteristics from DNA can provide investigative leads to find unknown perpetrators, and can also help in disaster victim and other missing person identification cases. Aiming for the application to both types of forensic casework, we previously developed and forensically validated the HIrisPlex test system enabling parallel DNA prediction of eye and hair colour. Although a recent proof-of-principle study demonstrated the general suitability of the HIrisPlex system for successfully analysing DNA from bones and teeth of various storage times and conditions, practical case applications to human remains are scarce. In this study, we applied the HIrisPlex system to 49 DNA samples obtained from bones or teeth of World War II victims excavated at six sites, mostly mass graves, in Slovenia. PCR-based DNA quantification ranged from 4 pg/μl to 313 pg/μl and on an average was 41 pg/μl across all samples. All 49 samples generated complete HIrisPlex profiles with the exception of one MC1R DNA marker (N29insA) missing in 83.7% of the samples. In 44 of the 49 samples (89.8%) complete 15-loci autosomal STR (plus amelogenin) profiles were obtained. Of 5 pairs of skeletal remains for which STR profiling suggested an origin in the same individuals, respectively, 4 showed the same HIrisPlex profiles and predicted eye and hair colours, respectively, while discrepancies in one pair (sample 26 and 43) are likely to be explained by DNA quantity and quality issues observed in sample 43. Sample 43 had the lowest DNA concentration of only 4 pg/μl, producing least reliable STR results and could be misleading in concluding that samples 43 and 26 originate from the same individual. The HIrisPlex-predicted eye and hair colours from two skeletal samples, suggested to derive from two brothers via STR profiling together with a living sister, were confirmed by the living sister's report. Overall, we demonstrate that after more than 70 years, HIrisPlex-based eye and hair colour prediction from skeletal remains is feasible with high success rate. Our results further encourage the use of the HIrisPlex system in missing person/disaster victim identification to aid the identification process in cases where ante-mortem samples or putative relatives are not directly available, and DNA predicted eye and hair colour information provides leads for locating them, allowing STRbased individual identification.
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- Epidemiology: DNA identifications after the 9/11 world trade center attack.Science. 2005; 310: 1122-1123
- International Society for Forensic Genetics (2007) DNA Commission of the International Society for Forensic Genetics (ISFG): recommendations regarding the role of forensic genetics for disaster victim identification (DVI).Forensic Sci. Int. Genet. 2007; 1: 3-12
- The contribution of DNA to the disaster victim identification (DVI) effort.Forensic Sci. Int. 2011; 205: 52-58
- DNA analysis in disaster victim identification.Forensic Sci. Med. Pathol. 2012; 8: 140-147
- Ancient DNA.Nat. Rev. Genet. 2001; 2: 353-359
- DNA preservation: a microsatellite-DNA study on ancient skeletal remains.Electrophoresis. 1999; 20: 1722-1728
- Teeth as a source of DNA for forensic identification of human remains:a review.Sci. Justice. 2013; 53: 433-441
- Genetic analyses from ancient DNA.Annu. Rev. Genet. 2004; 38: 645-679
- Assessing ancient DNA studies.Trends Ecol. Evol. 2005; 20: 541-544
- Typing short amplicon binary polymorphisms: supplementary SNP and Indel genetic information in the analysis of highly degraded skeletal remains.Forensic Sci. Int. Genet. 2012; 6: 469-476
- DNA in ancient bones −where is it located and how should we extract it.Ann. Anat. 2013; 194: 7-16
- Ancient human genome sequence of an extinct Palaeo-Eskimo.Nature. 2010; 463: 757-762
- New insights into the Tyrolean Iceman's origin and phenotype as inferred by whole-genome sequencing.Nat. Commun. 2012; 28: 698
- Re-inventing ancient human DNA.Investig. Genet. 2015; 6: 4
- Issues and strategies in the DNA identification of World Trade Center victims.Theor. Popul. Biol. 2003; 63: 173-178
- Bona fide colour: DNA prediction of human eye and hair colour from ancient and contemporary skeletal remains.Investig. Genet. 2013; 4: 3
- DNA identification by pedigree likelihood ratio accommodating population substructure and mutations.Investig. Genet. 2010; 1: 8
- Molecular genetic identification of skeletal remains from the Second world war Konfin I mass grave in Slovenia.Int. J. Legal Med. 2010; 124: 307-317
- Genetic Identification of Second World War Victim's Skeletal Remains.Lap Lambert Academic Publishing, Saarbrücken2013
- Example of human individual identification from World War II gravesite.Forensic Sci. Int. 2013; 233: 179-192
- Identification of missing norwegian World War II soldiers, in Karelia Russia.J. Forensic Sci. 2015; 60: 1104-1110
- Improving human forensics through advances in genetics, genomics and molecular biology.Nat. Rev. Genet. 2011; 12: 179-192
- Forensic DNA Phenotyping: predicting human appearance from crime scene material for investigative purposes.Forensic Sci. Int. Genet. 2015; 18: 33-48
- The HIrisPlex system for simultaneous prediction of hair and eye colour from DNA.Forensic Sci. Int. Genet. 2013; 7: 98-115
- Developmental validation of the HIrisPlex system: DNA-based eye and hair colour prediction for forensic and anthropological usage.Forensic Sci. Int. Genet. 2014; 9: 150-161
- Identification of the remains of King Richard III.Nat. Commun. 2014; 5: 5631
- Topografija evidentiranih grobišč (Topography of documented mass graves).in: Dežman J. Poročilo Komisije Vlade Republike Slovenije Za Reševanje Vprašanj Prikritih Grobišč 2005–2008. Družina, Ljubljana2008: 7-27
- Validation of mitochondrial DNA sequencing for forensic casework analysis.Int. J. Legal Med. 1995; 108: 68-74
- Automatable full demineralization DNA extraction procedure from degraded skeletal remains.Forensic Sci. Int. Genet. 2012; 6: 398-406
- Ancient DNA extraction from bones and teeth.Nat. Protoc. 2007; 2: 1756-1762
- A DNA extraction method of small quantities of bone for high-quality genotyping.Forensic Sci. Int. Genet. 2013; 7: 488-493
- DNA typing from skeletal remains: evaluation of multiplex and megaplex STR systems on DNA isolated from bone and teeth samples.Croat. Med. J. 2001; 42: 260-266
- DNA Commission of the International Society for Forensic Genetics: revised and extended guidelines for mitochondrial DNA typing.Forensic Sci. Int. Genet. 2014; 13: 134-142
- Highly efficient automated extraction of DNA from old and contemporary skeletal remains.J. Forensic Legal Med. 2016; 37: 78-86
- Extraction of DNA from human skeletal material.in: Forensic DNA Typing Protocols, Methods in Molecular Biology. vol 1420. Springer Science & Business Media, LLC, New York2016: 89-108
- EZ1 DNA Investigator Handbook.Hilden, 2014
- Quantifiler™ Human DNA Quantification kit User Guide.ThermoFisher Scientific, Foster City (CA)2003 (http://www3.appliedbiosystems.com/cms/groups/applied_markets_support/documents/generaldocuments/cms_041395.pdf)
- AmpFlSTR® NGMTM PCR amplification kit User Guide.ThermoFisher Scientific, Foster City (CA)2009 (https://www3.appliedbiosystems.com/cms/groups/applied_markets_support/documents/generaldocuments/cms_074250.pdf)
- Investigator ESSplex Plus Handbook.Hilden, 2013
- Highly efficient nuclear DNA typing of the World War II skeletal remains using three new autosomal short tandem repeat amplification kits with the extended European Standard Set of loci.Croat. Med. J. 2012; 53: 17-23
- Eye colour and the prediction of complex phenotypes from genotypes.Curr. Biol. 2009; 19: R192-3
- IrisPlex: a sensitive DNA tool for accurate prediction of blue and brown eye colour in the absence of ancestry information.Forensic Sci. Int. Genet. 2011; 5: 170-180
- DNA-based eye colour prediction across Europe with the IrisPlex system.Forensic Sci. Int. Genet. 2012; 6: 330-340
Published online: October 06, 2016
Accepted: October 6, 2016
Received in revised form: September 26, 2016
Received: March 24, 2016
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