title: Methylation Markers for the Identification of Body Fluids and Tissues from Forensic Trace Evidence.
abstract: The identification of body fluids is an essential tool for clarifying the course of events at a criminal site. The analytical problem is the fact that the biological material has been very often exposed to detrimental exogenous influences. Thereby, the molecular substrates used for the identification of the traces may become degraded. So far, most protocols utilize cell specific proteins or RNAs. Instead of measuring these more sensitive compounds this paper describes the application of the differential DNA-methylation. As a result of two genome wide screenings with the Illumina HumanMethylation BeadChips 27 and 450k we identified 150 candidate loci revealing differential methylation with regard to the body fluids venous blood, menstrual blood, vaginal fluid, saliva and sperm. Among them we selected 9 loci as the most promising markers. For the final determination of the methylation degree we applied the SNuPE-method. Because the degree of methylation might be modified by various endogenous and exogenous factors, we tested each marker with approximately 100 samples of each target fluid in a validation study. The stability of the detection procedure is proved in various simulated forensic surroundings according to standardized conditions. We studied the potential influence of 12 relatively common tumors on the methylation of the 9 markers. For this purpose the target fluids of 34 patients have been analysed. Only the cervix carcinoma might have an remarkable effect because impairing the signal of both vaginal markers. Using the Illumina MiSeq device we tested the potential influence of cis acting sequence variants on the methylation degree of the 9 markers in the specific body fluid DNA of 50 individuals. For 4 marker loci we observed such an influence either by sole SNPs or haplotypes. The identification of each target fluid is possible in arbitrary mixtures with the remaining four body fluids. The sensitivity of the individual body fluid tests is in the same range as for the forensic STR-analysis. It is the first forensic body fluid protocol which considers the exogenic and endogenic parameters potentially interfering with the true results.
authors.affiliation: Labor für Abstammungsbegutachtungen GmbH, Rheinbach, Germany.
authors.forename: Sophia
authors.initials: S
authors.lastname: Forat
authors.affiliation: Max Planck Genome Centre Cologne Institute for Breeding Research, Cologne, Germany.
authors.forename: Bruno
authors.initials: B
authors.lastname: Huettel
authors.affiliation: Max Planck Genome Centre Cologne Institute for Breeding Research, Cologne, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Reinhardt
authors.affiliation: Institute for Medical Biometry, Informatics and Epidemiology, University of Bonn, Bonn, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Fimmers
authors.affiliation: Department of Dermatology, Andrology Unit, University of Bonn, Bonn, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Haidl
authors.affiliation: Department of OB/GYN Hochtaunuskliniken Bad Homburg, Bad Homburg, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Denschlag
authors.affiliation: Labor für Abstammungsbegutachtungen GmbH, Rheinbach, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Olek
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D001826:Body Fluids / Q000378:metabolism*; D003198:Computer Simulation; D019175:DNA Methylation / Q000235:genetics*; D016002:Discriminant Analysis; D005260:Female; D053803:Forensic Genetics; D056426:Genetic Loci; D015894:Genome, Human; D006801:Humans; D008297:Male; D009928:Organ Specificity; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers
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