A data-driven, high-throughput methodology to determine tissue-specific differentially methylated regions able to discriminate body fluids

Joana Antunes, Quentin Gauthier, Vanessa Aguiar-Pulido, George Duncan, Bruce McCord

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Tissue-specific differentially methylated regions (tDMRs) are regions of the genome with methylation patterns that modulate gene expression in those tissue types. The detection of tDMRs in forensic evidence can permit the identification of body fluids at trace levels. In this report, we have performed a bioinformatic analysis of an existing array dataset to determine if new tDMRs could be identified for use in body fluid identification from forensic evidence. Once these sites were identified, primers were designed and bisulfite modification was performed. The relative methylation level for each body fluid at a given locus was then determined using qPCR with high-resolution melt analysis (HRM). After screening 127 tDMR's in multiple body fluids, we were able to identify four new markers able to discriminate blood (2 markers), vaginal epithelia (1 marker) and buccal cells (1 marker). One marker for each target body fluid was also tested with pyrosequencing showing results consistent with those obtained by HRM. This work successfully demonstrates the ability of in silico analysis to develop a novel set of tDMRs capable of being differentiated by real time PCR/HRM. The method can rapidly determine the body fluids left at crime scenes, assisting the triers of fact in forensic casework.

Original languageEnglish (US)
Pages (from-to)1168-1176
Number of pages9
JournalELECTROPHORESIS
Volume42
Issue number9-10
DOIs
StatePublished - May 2021

Keywords

  • Body fluid identification
  • DNA methylation
  • Forensic epigenetics
  • Infinium HumanMethylation450 BeadChip array
  • Serology

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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