An assessment of a massively parallel sequencing approach for the identification of individuals from mass graves of the Spanish Civil War (1936–1939)

Next‐generation sequencing technologies have opened new opportunities in forensic genetics. Here, we assess the applicability and performance of the MiSeq FGx™ & ForenSeq™ DNA Signature Prep Kit (Illumina) for the identification of individuals from the mass graves of the Spanish Civil War (1936–1939). The main limitations for individual identification are the low number of possible first‐degree living relatives and the high levels of DNA degradation reported in previous studies. Massively parallel sequencing technologies enabling the analysis of hundreds of regions and prioritizing short length amplicons constitute a promising tool for this kind of approaches. In this study, we first explore the power of this new technology to detect first‐ and second‐degree kinship given different scenarios of DNA degradation. Second, we specifically assess its performance in a set of low DNA input samples previously analyzed with CE technologies. We conclude that this methodology will allow identification of up to second‐degree relatives, even in situations with low sequencing performance and important levels of allele drop‐out; it is thus a technology that resolves previous drawbacks and that will allow a successful approximation to the identification of remains.

[1]  Jaume Bertranpetit,et al.  Human Genetic Variation, Shared and Private , 2012, Science.

[2]  Niels Morling,et al.  Second-generation sequencing of forensic STRs using the Ion Torrent™ HID STR 10-plex and the Ion PGM™. , 2015, Forensic science international. Genetics.

[3]  Francesc Calafell,et al.  Y-chromosome diversity in Catalan surname samples: insights into surname origin and frequency , 2015, European Journal of Human Genetics.

[4]  Bruce Budowle,et al.  Analysis of DNA forensic markers using high throughput mass spectrometry , 2009 .

[5]  Jaume Bertranpetit,et al.  A Melanocortin 1 Receptor Allele Suggests Varying Pigmentation Among Neanderthals , 2007, Science.

[6]  B. Bayer,et al.  Quantifiler Human DNA Quantification Kit (Applied Biosystems) as a screening kit for DNA profiling , 2009 .

[7]  Bruce Budowle,et al.  Evaluation of the Illumina(®) Beta Version ForenSeq™ DNA Signature Prep Kit for use in genetic profiling. , 2016, Forensic science international. Genetics.

[8]  C. Thermes,et al.  Ten years of next-generation sequencing technology. , 2014, Trends in genetics : TIG.

[9]  Niklas Söderholm,et al.  Repatriation and identification of the Finnish World War II soldiers. , 2007, Croatian medical journal.

[10]  Jože Balažic,et al.  Molecular genetic identification of skeletal remains from the Second World War Konfin I mass grave in Slovenia , 2010, International Journal of Legal Medicine.

[11]  M. V. Lareu,et al.  Global population variability in Promega PowerPlex CS7, D6S1043, and Penta B STRs , 2013, International Journal of Legal Medicine.

[12]  H. Poinar The top 10 list: criteria of authenticity for DNA from ancient and forensic samples , 2003 .

[13]  Herbert Oberacher,et al.  Increasing the discrimination power of forensic STR testing by employing high-performance mass spectrometry, as illustrated in indigenous South African and Central Asian populations , 2010, International Journal of Legal Medicine.

[14]  Yaran Yang,et al.  Application of Next-generation Sequencing Technology in Forensic Science , 2014, Genom. Proteom. Bioinform..

[15]  K. Bentayebi,et al.  Genetic analysis of 12 X-chromosome STRs in Western Mediterranean populations , 2015, International Journal of Legal Medicine.

[16]  Herbert Oberacher,et al.  Increased forensic efficiency of DNA fingerprints through simultaneous resolution of length and nucleotide variability by high‐performance mass spectrometry , 2008, Human mutation.

[17]  Sergio Cardoso,et al.  Digging up the recent Spanish memory: genetic identification of human remains from mass graves of the Spanish Civil War and posterior dictatorship. , 2015, Forensic science international. Genetics.

[18]  Bruce Budowle,et al.  Enhancing resolution and statistical power by utilizing mass spectrometry for detection of SNPs within the short tandem repeats , 2009 .

[19]  P. J. van der Zaag,et al.  Targeted enrichment of genomic DNA regions for next-generation sequencing , 2011, Briefings in functional genomics.

[20]  M. Subirana,et al.  La antropología forense al servicio de la justicia y la historia: las fosas de la Guerra Civil , 2010 .