RT Journal Article
T1 Development and optimization of the VISAGE basic prototype tool for forensic age estimation
A1 Heidegger, A.
A1 Xavier, C.
A1 Niederstätter, H.
A1 Puente Vila, María del Carmen de la
A1 Pospiech, E.
A1 Pisarek, A.
A1 Kayser, M.
A1 Branicki, W.
A1 Parson, W.
K1 Bisulfite PCR multiplex development
K1 Targeted bisulfite sequencing
K1 Age estimation
K1 MiSeq FGx sequencing
AB The VISAGE (VISible Attributes through GEnomics) consortium aims to develop, optimize and validate prototype tools to broaden the use of DNA intelligence methods in forensic routine laboratories. This includes age estimation based on the quantification of DNA methylation at specific CpG sites. Here, we present the VISAGE basic prototype tool for age estimation targeting 32 CpGs from five genes ELOVL2, MIR29B2CHG (herein, MIR29B2C), FHL2, TRIM59 and KLF14. The assay interrogates these well described age markers by multiplex PCR for bisulfite converted DNA and massively parallel sequencing on a MiSeq FGx instrument. We describe protocol optimizations including tests on five bisulfite conversion kits and an evaluation of the assay’s reproducibility and sensitivity with artificially methylated DNA standards. We observed robust quantification of methylation levels with a mean standard deviation of 1.4 % across ratios. Sensitivity tests showed no increase of variability down to 20 ng DNA input into bisulfite conversion with a median difference below 1.6 % between technical replicates
PB Elsevier
YR 2020
FD 2020-06-06
LK https://hdl.handle.net/10347/45546
UL https://hdl.handle.net/10347/45546
LA eng
NO A. Heidegger, C. Xavier, H. Niederstätter, M. de la Puente, E. Pośpiech, A. Pisarek, M. Kayser, W. Branicki, W. Parson, Development and optimization of the VISAGE basic prototype tool for forensic age estimation, Forensic Science International: Genetics, Volume 48, 2020, 102322, ISSN 1872-4973, https://doi.org/10.1016/j.fsigen.2020.102322
NO The study received support from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 740580 within the framework of the VISible Attributes through GEnomics (VISAGE) Project and Consortium. MdlP is supported by a postdoctoral fellowship awarded by the Consellería de Cultura, Educación e Ordenación Universitaria and the Consellería de Economía, Emprego e Industria from Xunta de Galicia (Modalidade A, ED481B 2017/088). We would like to thank Martin Steinlechner and Burkhard Berger for their help with blood sampling and Mayra Eduardoff for laboratory support (all Institute of Legal Medicine, Medical University of Innsbruck)
DS Minerva
RD 27 abr 2026