Novel identification of biofluids using a multiplex methylation sensitive restriction enzyme-PCR system

作者：Lin Yu Chih; Tsai Li Chin; Lee James Chun I; Su Chih Wen; Tzen Jason Tze Cheng; Linacre Adrian; Hsieh Hsing Mei^*

来源：Forensic Science International-Genetics, 2016, 25: 157-165.

DOI：10.1016/j.fsigen.2016.08.011

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

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 (R) MiniFiler (TM) 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.

• 出版日期2016-11