ABSTRACT
DNA methylation is known as a biomarker for age with applications in forensics. Here we describe the VISAGE (VISible Attributes through GEnomics) Consortium's enhanced tool for epigenetic age estimation in somatic tissues. The tool is based on eight DNA methylation markers (44 CpGs), bisulfite multiplex PCR followed by sequencing on the MiSeq FGx platform, and three statistical prediction models for blood, buccal cells and bones. The model for blood is based on six CpGs from ELOVL2, MIR29B2CHG, KLF14, FHL2, TRIM59 and PDE4C, and predicts age with a mean absolute error (MAE) of 3.2 years, while the model for buccal cells includes five CpGs from PDE4C, MIR29B2CHG, ELOVL2, KLF14 and EDARADD and predicts age with MAE of 3.7 years, and the model for bones has six CpGs from ELOVL2, KLF14, PDE4C and ASPA and predicts age with MAE of 3.4 years. The VISAGE enhanced tool for age estimation in somatic tissues enables reliable collection of DNA methylation data from small amounts of DNA using a sensitive multiplex MPS assay that provides accurate estimation of age in blood, buccal swabs, and bones using the statistical model tailored to each tissue.
Subject(s)
Aging/genetics , CpG Islands , Models, Statistical , Adolescent , Adult , Aged , Aged, 80 and over , Amidohydrolases/genetics , Blood Chemical Analysis , Bone and Bones/chemistry , Child , Child, Preschool , Cyclic Nucleotide Phosphodiesterases, Type 4/genetics , DNA Methylation , Edar-Associated Death Domain Protein/genetics , Epigenesis, Genetic , Fatty Acid Elongases/genetics , Female , High-Throughput Nucleotide Sequencing , Humans , Infant , Kruppel-Like Transcription Factors/genetics , Male , Middle Aged , Mouth Mucosa/chemistry , Multiplex Polymerase Chain Reaction , Sequence Analysis, DNA , Young AdultABSTRACT
Forensic DNA phenotyping is gaining interest as the number of applications increases within the forensic genetics community. The possibility of providing investigative leads in addition to conventional DNA profiling for human identification provides new insights into otherwise "cold" police investigations. The ability of reporting on the bio-geographical ancestry (BGA), appearance characteristics and age based on DNA obtained from a crime scene sample of an unknown donor makes the exploration of such markers and the development of new methods meaningful for criminal investigations. The VISible Attributes through GEnomics (VISAGE) Consortium aims to disseminate and broaden the use of predictive markers and develop fully optimized and validated prototypes for forensic casework implementation. Here, the first VISAGE appearance and ancestry tool development, performance and validation is reported. A total of 153 SNPs (96.84 % assay conversion rate) were successfully incorporated into a single multiplex reaction using the AmpliSeq™ design pipeline, and applied for massively parallel sequencing with the Ion S5 platform. A collaborative effort involving six VISAGE laboratory partners was devised to perform all validation tests. An extensive validation plan was carefully organized to explore the assay's overall performance with optimum and low-input samples, as well as with challenging and casework mock samples. In addition, forensic validation studies such as concordance and mixture tests recurring to the Coriell sample set with known genotypes were performed. Finally, inhibitor tolerance and specificity were also evaluated. Results showed a robust, highly sensitive assay with good overall concordance between laboratories.
Subject(s)
DNA Fingerprinting , DNA/genetics , High-Throughput Nucleotide Sequencing , Polymorphism, Single Nucleotide , Racial Groups/genetics , Software , Genetic Markers , Humans , Phenotype , Polymerase Chain Reaction , Reproducibility of Results , Sequence Analysis, DNAABSTRACT
Haplotype and allele frequencies for 17 Y-STR loci (DYS456, DYS389I/II, DYS390, DYS458, DYS19, DYS385 I/II, Y GATA H4, DYS437, DYS438, DYS448, DYS393, DYS391, DYS439, Y GATA C4, DYS392) were determined in 255 unrelated males from central Poland using AmpFlSTR Yfiler PCR Amplification Kit. Two hundred and fifty-two different haplotypes were observed. The most common three haplotypes were shared by 0.8% of the sample, respectively. Two hundred and forty-nine haplotypes were encountered only once.
Subject(s)
Chromosomes, Human, Y , Genetics, Population , Haplotypes , Tandem Repeat Sequences , DNA Fingerprinting , Gene Frequency , Humans , Male , Poland , Polymerase Chain ReactionABSTRACT
Allele frequencies for the fifteen STRs included in the GenePrint PowerPlex kit were determined in 870 unrelated individuals from Polish population. All loci met Hardy-Weinberg expectations. Exact tests disequilibrium analysis revealed no departures from independence out of 105 pair-wise comparisons. The combined MP and PE for all 15 loci are 6.00x10(-17) and 0.999999, respectively.
