Title

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

Authors

Joana Antunes, Florida International University
Quentin Gauthier, Florida International University
Vanessa Aguiar-Pulido, University of Miami
George Duncan, Nova Southeastern UniversityFollow
Bruce McCord, Florida International University

Document Type

Article

Publication Date

2-11-2021

Publication Title

Electrophoresis

Keywords

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

ISSN

1522-2683

Volume

42

Issue/No.

9-10

First Page

1168

Last Page

1176

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.

Comments

National Institute of Justice. Grant Numbers: 2015-R2-CX-0012, 2017-NX-BX-0001

Additional Comments

© 2021 Wiley-VCH GmbH

NSUWorks Citation

Antunes, Joana; Quentin Gauthier; Vanessa Aguiar-Pulido; George Duncan; and Bruce McCord. 2021. "A data-driven, high-throughput methodology to determine tissue-specific differentially methylated regions able to discriminate body fluids." Electrophoresis 42, (9-10): 1168-1176. doi:10.1002/elps.202000217.

ORCID ID

0000-0002-4931-5316

DOI

10.1002/elps.202000217