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Sökning: WFRF:(Junker M) > (2020-2023) > Development and int...

  • Heidegger, A.Medical University of Innsbruck (författare)

Development and inter-laboratory validation of the VISAGE enhanced tool for age estimation from semen using quantitative DNA methylation analysis

  • Artikel/kapitelEngelska2022

Förlag, utgivningsår, omfång ...

  • Elsevier BV,2022

Nummerbeteckningar

  • LIBRIS-ID:oai:lup.lub.lu.se:7f095b4f-c90f-4e64-85f1-f230dd97d65a
  • https://lup.lub.lu.se/record/7f095b4f-c90f-4e64-85f1-f230dd97d65aURI
  • https://doi.org/10.1016/j.fsigen.2021.102596DOI

Kompletterande språkuppgifter

  • Språk:engelska
  • Sammanfattning på:engelska

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Klassifikation

  • Ämneskategori:art swepub-publicationtype
  • Ämneskategori:ref swepub-contenttype

Anmärkningar

  • The analysis of DNA methylation has become an established method for chronological age estimation. This has triggered interest in the forensic community to develop new methods for age estimation from biological crime scene material. Various assays are available for age estimation from somatic tissues, the majority from blood. Age prediction from semen requires different DNA methylation markers and the only assays currently developed for forensic analysis are based on SNaPshot or pyrosequencing. Here, we describe a new assay using massively parallel sequencing to analyse 13 candidate CpG sites targeted in two multiplex PCRs. The assay has been validated by five consortium laboratories of the VISible Attributes through GEnomics (VISAGE) project within a collaborative exercise and was tested for reproducible quantification of DNA methylation levels and sensitivity with DNA methylation controls. Furthermore, DNA extracts and stains on Whatman FTA cards from two semen samples were used to evaluate concordance and mimic casework samples. Overall, the assay yielded high read depths (> 1000 reads) at all 13 marker positions. The methylation values obtained indicated robust quantification with an average standard deviation of 2.8% at the expected methylation level of 50% across the 13 markers and a good performance with 50 ng DNA input into bisulfite conversion. The absolute difference of quantifications from one participating laboratory to the mean quantifications of concordance and semen stains of remaining laboratories was approximately 1%. These results demonstrated the assay to be robust and suitable for age estimation from semen in forensic investigations. In addition to the 13-marker assay, a more streamlined protocol combining only five age markers in one multiplex PCR was developed. Preliminary results showed no substantial differences in DNA methylation quantification between the two assays, indicating its applicability with the VISAGE age model for semen developed with data from the complete 13-marker tool.

Ämnesord och genrebeteckningar

Biuppslag (personer, institutioner, konferenser, titlar ...)

  • Pisarek, A.Jagiellonian University (författare)
  • de la Puente, M.Medical University of Innsbruck,CIBER de Fisiopatología Obesidad y Nutricion (CIBEROBN) (författare)
  • Niederstätter, H.Medical University of Innsbruck (författare)
  • Pośpiech, E.Jagiellonian University (författare)
  • Woźniak, A.Central Forensic Laboratory of the Police (författare)
  • Schury, N.Federal Criminal Police Office, Wiesbaden (författare)
  • Unterländer, M.Federal Criminal Police Office, Wiesbaden (författare)
  • Sidstedt, M.Swedish Police Authority(Swepub:lu)tmb-mas (författare)
  • Junker, K.Swedish Police Authority (författare)
  • Ventayol Garcia, M.Swedish Police Authority,Netherlands Forensic Institute (författare)
  • Laurent, F. X.Institut National de Police Scientifique (författare)
  • Ulus, A.Institut National de Police Scientifique (författare)
  • Vannier, J.Institut National de Police Scientifique (författare)
  • Bastisch, I.Federal Criminal Police Office, Wiesbaden (författare)
  • Hedman, J.Lund University,Lunds universitet,Teknisk mikrobiologi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Applied Microbiology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Swedish Police Authority(Swepub:lu)tmb-jeh (författare)
  • Sijen, T.University of Amsterdam,Netherlands Forensic Institute (författare)
  • Branicki, W.Jagiellonian University (författare)
  • Xavier, C.Medical University of Innsbruck (författare)
  • Parson, W.Pennsylvania State University,Medical University of Innsbruck (författare)
  • Medical University of InnsbruckJagiellonian University (creator_code:org_t)

Sammanhörande titlar

  • Ingår i:Forensic Science International: Genetics: Elsevier BV561872-4973

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