Novel Identification of Biofluids Using a Multiplex Methylation-specific PCR Combined with Single-base Extension System

Overview

Journal Forensic Sci Med Pathol

Specialties Forensic Sciences
Pathology

Date 2016 Mar 15

PMID 26972905

Citations 8

Authors

Yu-Chih Lin

Li-Chin Tsai

James Chun-I Lee

Kuo-Lan Liu

Jason Tze-Cheng Tzen

Adrian Linacre

Hsing-Mei Hsieh

Affiliations

Soon will be listed here.

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.

Citing Articles

Application of a new composite genetic marker semen-specific methylation-microhaplotype in the analysis of semen-vaginal fluid mixtures.

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Expanding the scope of methylation-sensitive restriction enzyme (MSRE) PCR for forensic identification of body fluids through the novel use of methylation-dependent restriction enzymes (MDRE) and the combination of autosomal and Y-chromosomal markers.

Rothe J, Becker J, Charchinezhadamouei M, Mahr S, Lembeck F, Dannemann N Int J Legal Med. 2023; 138(2):375-393.

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Development of a novel panel for blood identification based on blood-specific CpG-linked SNP markers.

Li Z, Liu N, Yuan F, Guan Z, Liu J, Liu F Int J Legal Med. 2023; 138(3):1205-1219.

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More than just blood, saliva, or sperm-setup of a workflow for body fluid identification by DNA methylation analysis.

Konrad H, Jurgens L, Hartung B, Poetsch M Int J Legal Med. 2023; 137(6):1683-1692.

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Knife wound or nosebleed-where does the blood at the crime scene come from?.

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PMID: 37148347 PMC: 10247842. DOI: 10.1007/s00414-023-03012-2.

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