Novel identification of biofluids using a multiplex methylation sensitive restriction enzyme-PCR system.
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.
Graduate Institute of Biotechnology, National Chung Hsing University, 250 Kuo Kuang Road, South District, Taichung City 40227, Taiwan, ROC; Taichung City Government Police Department, 588 Wenxin Road Section 2, Xitun District, Taichung City 40758, Taiwan, ROC.
Yu-Chih
YC
Lin
Department of Forensic Science, Central Police University, 56 Shu-Jen Road, Kwei-San, Taoyuan 33304, Taiwan, ROC.
Li-Chin
LC
Tsai
Department of Forensic Medicine, College of Medicine, National Taiwan University, 1 Jen-Ai Road Section 1, Taipei 10051, Taiwan, ROC.
James Chun-I
JC
Lee
Department of Forensic Science, Central Police University, 56 Shu-Jen Road, Kwei-San, Taoyuan 33304, Taiwan, ROC; Forensic Biology Division, Criminal Investigation Bureau, National Police Agency, 5 Lane 553, Zhongxiao East Road Section 4, Xinyi District, Taipei 11072, Taiwan, ROC.
Chih-Wen
CW
Su
Graduate Institute of Biotechnology, National Chung Hsing University, 250 Kuo Kuang Road, South District, Taichung City 40227, Taiwan, ROC.
Jason Tze-Cheng
JT
Tzen
School of Biological Sciences, Flinders University, Adelaide 5001, Australia.
Adrian
A
Linacre
Department of Forensic Science, Central Police University, 56 Shu-Jen Road, Kwei-San, Taoyuan 33304, Taiwan, ROC. Electronic address: mei@mail.cpu.edu.tw.
Hsing-Mei
HM
Hsieh
D001774:Blood Chemical Analysis; D002582:Cervix Mucus / Q000737:chemistry; D018899:CpG Islands / Q000235:genetics; D004247:DNA / Q000302:isolation & purification; D016172:DNA Fingerprinting; D019175:DNA Methylation; D004262:DNA Restriction Enzymes; D005260:Female; D005819:Genetic Markers; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D060885:Multiplex Polymerase Chain Reaction; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry; D012661:Semen / Q000737:chemistry
D005819:Genetic Markers; D004247:DNA; D004262:DNA Restriction Enzymes
Biofluid identification; Biofluid-specific methylated marker; Methylation sensitive restriction enzyme-PCR (MSRE-PCR); miniSTR