Independent validation of body fluid-specific CpG markers and construction of a robust multiplex assay

Hannah Holtkötter, Vanessa Beyer, Kristina Schwender, Alina Glaub, Kristina Schulze Johann, Marianne Schürenkamp, Ursula Sibbing, Sabrina Banken, Peter Wiegand, Heidi Pfeiffer, Lynn Dennany, Marielle Vennamann

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Potential forensic use of tissue-specific DNA methylation markers has recently been discussed for the identification of the biological source of a stain. In this study 13 promising markers were evaluated to identify suitable candidate markers for the development of a robust and reliable multiplex assay. The results of this study suggest that a combination of only four highly informative markers will be enough for clear body fluid identification. A multiplex assay was developed for the identification of menstrual blood, saliva, semen, and venous blood. This assay was successfully applied to the identification of these body fluids in mixtures and crime scene stains. The multiplex assay aids in the identification of not only single source body fluids but also of body fluid mixtures. The main advantage of using DNA methylation assays over alternative tests is that it can be applied at a later time point in the investigative process since testing is possible even after DNA analysis.
Original languageEnglish
Pages (from-to)261-268
Number of pages8
JournalForensic Science International: Genetics
Early online date9 May 2017
Publication statusPublished - 31 Jul 2017


  • body fluids
  • epigenetics markers
  • DNA methylation
  • multiplex
  • body fluid identification
  • crime scene stains
  • biological source


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