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

Holtkötter, Hannah and Beyer, Vanessa and Schwender, Kristina and Glaub, Alina and Schulze Johann, Kristina and Schürenkamp, Marianne and Sibbing, Ursula and Banken, Sabrina and Wiegand, Peter and Pfeiffer, Heidi and Dennany, Lynn and Vennamann, Marielle (2017) Independent validation of body fluid-specific CpG markers and construction of a robust multiplex assay. Forensic Science International: Genetics, 29. pp. 261-268. ISSN 1872-4973 (

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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.


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