TY - JOUR
T1 - Novel Identification of Biofluids Using a Multiplex Methylation Sensitive Restriction Enzyme-PCR System
AU - Lin, Yu-Chih
AU - Tsai, Li-Chin
AU - Lee, James
AU - Su, Chih-Wen
AU - Tzen, Jason Tze-Cheng
AU - Linacre, Adrian
AU - Hsieh, Hsing-Mei
PY - 2016/11/1
Y1 - 2016/11/1
N2 - 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.
AB - 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.
KW - Biofluid identification
KW - Biofluid-specific methylated marker
KW - Methylation sensitive restriction enzyme-PCR (MSRE-PCR)
KW - miniSTR
UR - http://www.scopus.com/inward/record.url?scp=84985920064&partnerID=8YFLogxK
U2 - 10.1016/j.fsigen.2016.08.011
DO - 10.1016/j.fsigen.2016.08.011
M3 - Article
SN - 1872-4973
VL - 25
SP - 157
EP - 165
JO - Forensic Science International: Genetics
JF - Forensic Science International: Genetics
ER -