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Forensic Science, Medicine and Pathology
Published in: Forensic Science, Medicine and Pathology 2/2016

01-06-2016 | Original Article

Novel identification of biofluids using a multiplex methylation-specific PCR combined with single-base extension system

Authors: Yu-Chih Lin, Li-Chin Tsai, James Chun-I Lee, Kuo-Lan Liu, Jason Tze-Cheng Tzen, Adrian Linacre, Hsing-Mei Hsieh

Published in: Forensic Science, Medicine and Pathology | Issue 2/2016

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Abstract

Purpose

Knowledge of the composition of complex body fluid mixtures may aid forensic investigations greatly. However, many of the traditional tests are presumptive in nature and can lead to ambiguous results. The aim of this study is to establish a reliable method to identify various biofluids via analysis of their DNA methylation profiles.

Methods

A total of eight biofluid-specific methylated markers for saliva, venous blood, vaginal fluids, and semen were isolated from the open database of Infinium HumanMethylation450 BeadChip. These biofluid-specific markers, a control marker to confirm bisulfite conversion, and a gender marker, were combined into a 10-plex methylation-specific PCR single-base-extension (MSP-SBE) system.

Results

Analysis of 65 DNA samples isolated from venous blood, semen, vaginal fluid, saliva, and menstrual blood that had been treated with bisulfite, resulted in all eight markers detecting the body fluid to which they were designed. Unambiguous body fluid identification occurred from both single sources of body fluids and complex mixtures. A threshold was devised for each marker to minimize the chance of a false inclusion. The efficacy of the assay and application to forensic practice was demonstrated using five non-probative samples from real alleged sexual assault cases. The system unambiguously determined the biofluid types for the non-probative forensic samples that previously resulted in inconclusive or conflicting results using traditional tests.

Conclusions

The results demonstrated the 10-plex MSP-SBE system established in this study is both sensitive and specific when applied to body fluid identification and can be readily adopted into forensic practice.
Appendix
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Metadata
Title
Novel identification of biofluids using a multiplex methylation-specific PCR combined with single-base extension system
Authors
Yu-Chih Lin
Li-Chin Tsai
James Chun-I Lee
Kuo-Lan Liu
Jason Tze-Cheng Tzen
Adrian Linacre
Hsing-Mei Hsieh
Publication date
01-06-2016
Publisher
Springer US
Published in
Forensic Science, Medicine and Pathology / Issue 2/2016
Print ISSN: 1547-769X
Electronic ISSN: 1556-2891
DOI
https://doi.org/10.1007/s12024-016-9763-3

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