Novel Identification of Biofluids Using a Multiplex Methylation Sensitive Restriction Enzyme-PCR System

Yu-Chih Lin, Li-Chin Tsai, James Lee, Chih-Wen Su, Jason Tze-Cheng Tzen, Adrian Linacre, Hsing-Mei Hsieh

    Research output: Contribution to journalArticlepeer-review

    21 Citations (Scopus)


    The identification of a specific body fluid encountered in a forensic investigation can give crucial information. This identification can be aided by methylation profiles based on selected markers specific to a range of biofluids. In this study, the open database of Infinium HumanMethylation450 BeadChip was searched for markers specific for semen, vaginal fluids, saliva, venous blood and menstrual blood. A total of 8 biofluid-specific methylated markers and 2 control markers were combined into a 10-plex methylation sensitive restriction enzyme-PCR (MSRE-PCR) system. Based upon the analysis of 100 DNA samples from these 5 biofluid types, unambiguous results were obtained to identify the body fluid from which it originated. Validation studies of the developed 10-plex MSRE-PCR included sensitivity, reproducibility and mixed body fluids. Co-amplification of the established MSRE-PCR system and the microsatellite loci in AmpFlSTR® MiniFiler™ PCR Amplification Kit was performed to generate both the methylation profile for biofluid type and the miniSTR profile. This allowed human identification and the identification of the body fluid type to be performed in a single reaction. The results of this study displayed the applicability of this 10-plex MSRE-PCR system in forensic science.

    Original languageEnglish
    Pages (from-to)157-165
    Number of pages9
    JournalForensic Science International: Genetics
    Publication statusPublished - 1 Nov 2016


    • Biofluid identification
    • Biofluid-specific methylated marker
    • Methylation sensitive restriction enzyme-PCR (MSRE-PCR)
    • miniSTR


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