title: Inter-laboratory evaluation of the EUROFORGEN Global ancestry-informative SNP panel by massively parallel sequencing using the Ion PGM™.
abstract: The EUROFORGEN Global ancestry-informative SNP (AIM-SNPs) panel is a forensic multiplex of 128 markers designed to differentiate an individual's ancestry from amongst the five continental population groups of Africa, Europe, East Asia, Native America, and Oceania. A custom multiplex of AmpliSeq™ PCR primers was designed for the Global AIM-SNPs to perform massively parallel sequencing using the Ion PGM™ system. This study assessed individual SNP genotyping precision using the Ion PGM™, the forensic sensitivity of the multiplex using dilution series, degraded DNA plus simple mixtures, and the ancestry differentiation power of the final panel design, which required substitution of three original ancestry-informative SNPs with alternatives. Fourteen populations that had not been previously analyzed were genotyped using the custom multiplex and these studies allowed assessment of genotyping performance by comparison of data across five laboratories. Results indicate a low level of genotyping error can still occur from sequence misalignment caused by homopolymeric tracts close to the target SNP, despite careful scrutiny of candidate SNPs at the design stage. Such sequence misalignment required the exclusion of component SNP rs2080161 from the Global AIM-SNPs panel. However, the overall genotyping precision and sensitivity of this custom multiplex indicates the Ion PGM™ assay for the Global AIM-SNPs is highly suitable for forensic ancestry analysis with massively parallel sequencing.
authors.affiliation: Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: M
authors.initials: M
authors.lastname: Eduardoff
authors.affiliation: Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Cologne, Germany.
authors.forename: T E
authors.initials: TE
authors.lastname: Gross
authors.affiliation: Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Santos
authors.affiliation: Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: M
authors.initials: M
authors.lastname: de la Puente
authors.affiliation: Faculty of Life Sciences and Medicine, King's College, London, UK.
authors.forename: D
authors.initials: D
authors.lastname: Ballard
authors.affiliation: Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: C
authors.initials: C
authors.lastname: Strobl
authors.affiliation: Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Børsting
authors.affiliation: Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: N
authors.initials: N
authors.lastname: Morling
authors.affiliation: Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: L
authors.initials: L
authors.lastname: Fusco
authors.affiliation: Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Hussing
authors.affiliation: Department of Genetics, Faculty of Science, Eötvös Loránd University Budapest, Hungary.
authors.forename: B
authors.initials: B
authors.lastname: Egyed
authors.affiliation: Department of Biology, University of Aveiro, Aveiro, Portugal.
authors.forename: L
authors.initials: L
authors.lastname: Souto
authors.affiliation: Fiji Police Forensic Biology and DNA Laboratory, Nasova, Suva, Fiji.
authors.forename: J
authors.initials: J
authors.lastname: Uacyisrael
authors.affiliation: Faculty of Life Sciences and Medicine, King's College, London, UK.
authors.forename: D
authors.initials: D
authors.lastname: Syndercombe Court
authors.affiliation: Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain; Center of Excellence in Genomic Medicine Research, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Á
authors.initials: Á
authors.lastname: Carracedo
authors.affiliation: Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: M V
authors.initials: MV
authors.lastname: Lareu
authors.affiliation: Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Cologne, Germany.
authors.forename: P M
authors.initials: PM
authors.lastname: Schneider
authors.affiliation: Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria; Forensic Science Program, The Pennsylvania State University, PA, USA.
authors.forename: W
authors.initials: W
authors.lastname: Parson
authors.affiliation: Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain. Electronic address: c.phillips@mac.com.
authors.forename: C
authors.initials: C
authors.lastname: Phillips
authors.affiliation: Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria; Forensic Science Program, The Pennsylvania State University, PA, USA.
authors.forename: W
authors.initials: W
authors.lastname: Parson
authors.affiliation: Forensic Genetics Unit, Institute of Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain. Electronic address: c.phillips@mac.com.
authors.forename: C
authors.initials: C
authors.lastname: Phillips
mesh_terms: D053836:DNA Degradation, Necrotic; D016172:DNA Fingerprinting; D017931:DNA Primers; D030541:Databases, Genetic; D005787:Gene Frequency; D005819:Genetic Markers; D005828:Genetics, Population; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation*; D006801:Humans; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D044469:Racial Groups / Q000235:genetics*
chemical_list: D017931:DNA Primers; D005819:Genetic Markers
keywords: Ancestry-informative SNPs; Forensic ancestry analysis; Ion PGM™; Massively parallel sequencing (MPS)