Skip to main content
Top
Published in: International Journal of Legal Medicine 1/2012

01-01-2012 | Original Article

Potential forensic application of DNA methylation profiling to body fluid identification

Authors: Hwan Young Lee, Myung Jin Park, Ajin Choi, Ja Hyun An, Woo Ick Yang, Kyoung-Jin Shin

Published in: International Journal of Legal Medicine | Issue 1/2012

Login to get access

Abstract

DNA analysis of various body fluid stains at crime scenes facilitates the identification of individuals but does not currently determine the type and origin of the biological material. Recent advances in whole genome epigenetic analysis indicate that chromosome pieces called tDMRs (tissue-specific differentially methylated regions) show different DNA methylation profiles according to the type of cell or tissue. We examined the potential of tissue-specific differential DNA methylation for body fluid identification. Five tDMRs for the genes DACT1, USP49, HOXA4, PFN3, and PRMT2 were selected, and DNA methylation profiles for these tDMRs were produced by bisulfite sequencing using pooled DNA from blood, saliva, semen, menstrual blood, and vaginal fluid. The tDMRs for DACT1 and USP49 showed semen-specific hypomethylation, and the tDMRs for HOXA4, PFN3, and PRMT2 displayed varying degrees of methylation according to the type of body fluid. Preliminary tests using methylation-specific PCR for the DACT1 and USP49 tDMRs showed that these two markers could be used successfully to identify semen samples including sperm cells. Body fluid-specific differential DNA methylation may be a promising indicator for body fluid identification. Because DNA methylation profiling uses the same biological source of DNA for individual identification profiling, the determination of more body fluid-specific tDMRs and the development of convenient tDMR analysis methods will facilitate the broad implementation of body fluid identification in forensic casework.
Appendix
Available only for authorised users
Literature
1.
go back to reference Virkler K, Lednev IK (2009) Analysis of body fluids for forensic purposes: from laboratory testing to non-destructive rapid confirmatory identification at a crime scene. Forensic Sci Int 188:1–17PubMedCrossRef Virkler K, Lednev IK (2009) Analysis of body fluids for forensic purposes: from laboratory testing to non-destructive rapid confirmatory identification at a crime scene. Forensic Sci Int 188:1–17PubMedCrossRef
2.
go back to reference Bauer M, Polzin S, Patzelt D (2003) Quantification of RNA degradation by semi-quantitative duplex and competitive RT-PCR: a possible indicator of the age of bloodstains? Forensic Sci Int 138:94–103PubMedCrossRef Bauer M, Polzin S, Patzelt D (2003) Quantification of RNA degradation by semi-quantitative duplex and competitive RT-PCR: a possible indicator of the age of bloodstains? Forensic Sci Int 138:94–103PubMedCrossRef
3.
go back to reference Bauer M, Patzelt D (2002) Evaluation of mRNA markers for the identification of menstrual blood. J Forensic Sci 47:1278–1282PubMed Bauer M, Patzelt D (2002) Evaluation of mRNA markers for the identification of menstrual blood. J Forensic Sci 47:1278–1282PubMed
4.
go back to reference Juusola J, Ballantyne J (2003) Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification. Forensic Sci Int 135:85–96PubMedCrossRef Juusola J, Ballantyne J (2003) Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification. Forensic Sci Int 135:85–96PubMedCrossRef
5.
go back to reference Juusola J, Ballantyne J (2005) Multiplex mRNA profiling for the identification of body fluids. Forensic Sci Int 152:1–12PubMedCrossRef Juusola J, Ballantyne J (2005) Multiplex mRNA profiling for the identification of body fluids. Forensic Sci Int 152:1–12PubMedCrossRef
6.
go back to reference Nussbaumer C, Gharehbaghi-Schnell E, Korschineck I (2006) Messenger RNA profiling: a novel method for body fluid identification by real-time PCR. Forensic Sci Int 157:181–186PubMedCrossRef Nussbaumer C, Gharehbaghi-Schnell E, Korschineck I (2006) Messenger RNA profiling: a novel method for body fluid identification by real-time PCR. Forensic Sci Int 157:181–186PubMedCrossRef
7.
go back to reference Juusola J, Ballantyne J (2007) mRNA profiling for body fluid identification by multiplex quantitative RT-PCR. J Forensic Sci 52:1252–1262PubMed Juusola J, Ballantyne J (2007) mRNA profiling for body fluid identification by multiplex quantitative RT-PCR. J Forensic Sci 52:1252–1262PubMed
8.
go back to reference Zubakov D, Hanekamp E, Kokshoorn M, van Ijcken W, Kayser M (2008) Stable RNA markers for identification of blood and saliva stains revealed from whole genome expression analysis of time-wise degraded samples. Int J Leg Med 122:135–142CrossRef Zubakov D, Hanekamp E, Kokshoorn M, van Ijcken W, Kayser M (2008) Stable RNA markers for identification of blood and saliva stains revealed from whole genome expression analysis of time-wise degraded samples. Int J Leg Med 122:135–142CrossRef
9.
go back to reference Hanson EK, Lubenow H, Ballantyne J (2009) Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs. Anal Biochem 387:303–314PubMedCrossRef Hanson EK, Lubenow H, Ballantyne J (2009) Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs. Anal Biochem 387:303–314PubMedCrossRef
10.
go back to reference Setzer M, Juusola J, Ballantyne J (2008) Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains. J Forensic Sci 53:296–305PubMedCrossRef Setzer M, Juusola J, Ballantyne J (2008) Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains. J Forensic Sci 53:296–305PubMedCrossRef
11.
go back to reference Zubakov D, Kokshoorn M, Kloosterman A, Kayser M (2009) New markers for old stains: stable mRNA markers for blood and saliva identification from up to 16-year-old stains. Int J Leg Med 123:71–74CrossRef Zubakov D, Kokshoorn M, Kloosterman A, Kayser M (2009) New markers for old stains: stable mRNA markers for blood and saliva identification from up to 16-year-old stains. Int J Leg Med 123:71–74CrossRef
12.
go back to reference Holliday R, Pugh JE (1975) DNA modification mechanisms and gene activity during development. Science 187:226–232PubMedCrossRef Holliday R, Pugh JE (1975) DNA modification mechanisms and gene activity during development. Science 187:226–232PubMedCrossRef
13.
14.
go back to reference Russo VEA, Martienssen RA, Riggs AD (1996) Epigenetic mechanisms of gene regulation. Cold Spring Harbor Laboratory Press, Plainview Russo VEA, Martienssen RA, Riggs AD (1996) Epigenetic mechanisms of gene regulation. Cold Spring Harbor Laboratory Press, Plainview
15.
go back to reference Tost J (ed) (2008) Epigenetics. Caister Academic Press, Norfolk Tost J (ed) (2008) Epigenetics. Caister Academic Press, Norfolk
16.
go back to reference Ohgane J, Yagi S, Shiota K (2008) Epigenetics: the DNA methylation profile of tissue-dependent and differentially methylated regions in cells. Placenta 29:S29–S35PubMedCrossRef Ohgane J, Yagi S, Shiota K (2008) Epigenetics: the DNA methylation profile of tissue-dependent and differentially methylated regions in cells. Placenta 29:S29–S35PubMedCrossRef
17.
go back to reference Christensen BC, Houseman EA, Marsit CJ, Zheng S, Wrensch MR, Wiemels JL, Nelson HH, Karagas MR, Padbury JF, Bueno R, Sugarbaker DJ, Yeh RF, Wiencke JK, Kelsey KT (2009) Aging and environmental exposures alter tissue-specific DNA methylation dependent upon CpG island context. PLoS Genet 5:e1000602PubMedCrossRef Christensen BC, Houseman EA, Marsit CJ, Zheng S, Wrensch MR, Wiemels JL, Nelson HH, Karagas MR, Padbury JF, Bueno R, Sugarbaker DJ, Yeh RF, Wiencke JK, Kelsey KT (2009) Aging and environmental exposures alter tissue-specific DNA methylation dependent upon CpG island context. PLoS Genet 5:e1000602PubMedCrossRef
18.
go back to reference Schilling E, Rehli M (2007) Global, comparative analysis of tissue-specific promoter CpG methylation. Genomics 90:314–323PubMedCrossRef Schilling E, Rehli M (2007) Global, comparative analysis of tissue-specific promoter CpG methylation. Genomics 90:314–323PubMedCrossRef
19.
go back to reference Kitamura E, Igarashi J, Morohashi A, Hida N, Oinuma T, Nemoto N, Song F, Ghosh S, Held WA, Yoshida-Noro C, Nagase H (2007) Analysis of tissue-specific differentially methylated regions (TDMs) in humans. Genomics 89:326–337PubMedCrossRef Kitamura E, Igarashi J, Morohashi A, Hida N, Oinuma T, Nemoto N, Song F, Ghosh S, Held WA, Yoshida-Noro C, Nagase H (2007) Analysis of tissue-specific differentially methylated regions (TDMs) in humans. Genomics 89:326–337PubMedCrossRef
20.
go back to reference Igarashi J, Muroi S, Kawashima H, Wang X, Shinojima Y, Kitamura E, Oinuma T, Nemoto N, Song F, Ghosh S, Held WA, Nagase H (2008) Quantitative analysis of human tissue-specific differences in methylation. Biochem Biophys Res Commun 376:658–664PubMedCrossRef Igarashi J, Muroi S, Kawashima H, Wang X, Shinojima Y, Kitamura E, Oinuma T, Nemoto N, Song F, Ghosh S, Held WA, Nagase H (2008) Quantitative analysis of human tissue-specific differences in methylation. Biochem Biophys Res Commun 376:658–664PubMedCrossRef
21.
go back to reference Song F, Mahmood S, Ghosh S, Liang P, Smiraglia DJ, Nagase H, Held WA (2009) Tissue specific differentially methylated regions (TDMR): changes in DNA methylation during development. Genomics 93:130–139PubMedCrossRef Song F, Mahmood S, Ghosh S, Liang P, Smiraglia DJ, Nagase H, Held WA (2009) Tissue specific differentially methylated regions (TDMR): changes in DNA methylation during development. Genomics 93:130–139PubMedCrossRef
22.
go back to reference Song F, Smith JF, Kimura MT, Morrow AD, Matsuyama T, Nagase H, Held WA (2005) Association of tissue-specific differentially methylated regions (TDMs) with differential gene expression. Proc Natl Acad Sci USA 102:3336–3341PubMedCrossRef Song F, Smith JF, Kimura MT, Morrow AD, Matsuyama T, Nagase H, Held WA (2005) Association of tissue-specific differentially methylated regions (TDMs) with differential gene expression. Proc Natl Acad Sci USA 102:3336–3341PubMedCrossRef
23.
go back to reference Illingworth R, Kerr A, Desousa D, Jørgensen H, Ellis P, Stalker J, Jackson D, Clee C, Plumb R, Rogers J, Humphray S, Cox T, Langford C, Bird A (2008) A novel CpG island set identifies tissue-specific methylation at developmental gene loci. PLoS Biol 6:e22PubMedCrossRef Illingworth R, Kerr A, Desousa D, Jørgensen H, Ellis P, Stalker J, Jackson D, Clee C, Plumb R, Rogers J, Humphray S, Cox T, Langford C, Bird A (2008) A novel CpG island set identifies tissue-specific methylation at developmental gene loci. PLoS Biol 6:e22PubMedCrossRef
24.
go back to reference Doi A, Park IH, Wen B, Murakami P, Aryee MJ, Irizarry R, Herb B, Ladd-Acosta C, Rho J, Loewer S, Miller J, Schlaeger T, Daley GQ, Feinberg AP (2009) Differential methylation of tissue- and cancer-specific CpG island shores distinguishes human induced pluripotent stem cells, embryonic stem cells and fibroblasts. Nat Genet 41:1350–1353PubMedCrossRef Doi A, Park IH, Wen B, Murakami P, Aryee MJ, Irizarry R, Herb B, Ladd-Acosta C, Rho J, Loewer S, Miller J, Schlaeger T, Daley GQ, Feinberg AP (2009) Differential methylation of tissue- and cancer-specific CpG island shores distinguishes human induced pluripotent stem cells, embryonic stem cells and fibroblasts. Nat Genet 41:1350–1353PubMedCrossRef
25.
go back to reference Li LC, Dahiya R (2002) MethPrimer: designing primers for methylation PCRs. Bioinformatics 18:1427–1431PubMedCrossRef Li LC, Dahiya R (2002) MethPrimer: designing primers for methylation PCRs. Bioinformatics 18:1427–1431PubMedCrossRef
26.
go back to reference Bock C, Reither S, Mikeska T, Paulsen M, Walter J, Lengauer T (2005) BiQ Analyzer: visualization and quality control for DNA methylation data from bisulfite sequencing. Bioinformatics 21:4067–4068PubMedCrossRef Bock C, Reither S, Mikeska T, Paulsen M, Walter J, Lengauer T (2005) BiQ Analyzer: visualization and quality control for DNA methylation data from bisulfite sequencing. Bioinformatics 21:4067–4068PubMedCrossRef
27.
go back to reference Rohde C, Zhang Y, Jurkowski TP, Stamerjohanns H, Reinhardt R, Jeltsch A (2008) Bisulfite sequencing Data Presentation and Compilation (BDPC) web server—a useful tool for DNA methylation analysis. Nucleic Acids Res 36:e34PubMedCrossRef Rohde C, Zhang Y, Jurkowski TP, Stamerjohanns H, Reinhardt R, Jeltsch A (2008) Bisulfite sequencing Data Presentation and Compilation (BDPC) web server—a useful tool for DNA methylation analysis. Nucleic Acids Res 36:e34PubMedCrossRef
28.
go back to reference Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386PubMed Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386PubMed
29.
go back to reference Bitu CC, De Souza Setubal Destro MF, Lopes MA, Jorge J, Graner E, Coletta RD (2008) Dysregulated expression of Hox genes in oral squamous cell carcinomas. Oral presentation #17 at 2008 International Association of Oral Pathologists, San Francisco, USA Bitu CC, De Souza Setubal Destro MF, Lopes MA, Jorge J, Graner E, Coletta RD (2008) Dysregulated expression of Hox genes in oral squamous cell carcinomas. Oral presentation #17 at 2008 International Association of Oral Pathologists, San Francisco, USA
30.
go back to reference Byun HM, Siegmund KD, Pan F, Weisenberger DJ, Kanel G, Laird PW, Yang AS (2009) Epigenetic profiling of somatic tissues from human autopsy specimens identifies tissue- and individual-specific DNA methylation patterns. Hum Mol Genet 18:4808–4817PubMedCrossRef Byun HM, Siegmund KD, Pan F, Weisenberger DJ, Kanel G, Laird PW, Yang AS (2009) Epigenetic profiling of somatic tissues from human autopsy specimens identifies tissue- and individual-specific DNA methylation patterns. Hum Mol Genet 18:4808–4817PubMedCrossRef
31.
go back to reference Sugimoto K, Koh E, Sin HS, Maeda Y, Narimoto K, Izumi K, Kobori Y, Kitamura E, Nagase H, Yoshida A, Namiki M (2009) Tissue-specific differentially methylated regions of the human VASA gene are potentially associated with maturation arrest phenotype in the testis. J Hum Genet 54:450–456PubMedCrossRef Sugimoto K, Koh E, Sin HS, Maeda Y, Narimoto K, Izumi K, Kobori Y, Kitamura E, Nagase H, Yoshida A, Namiki M (2009) Tissue-specific differentially methylated regions of the human VASA gene are potentially associated with maturation arrest phenotype in the testis. J Hum Genet 54:450–456PubMedCrossRef
33.
go back to reference Naito E, Dewa K, Fukuda M, Sumi H, Wakabayashi Y, Umetsu K, Yuasa I, Yamanouchi H (2003) Novel paternity testing by distinguishing parental alleles at a VNTR locus in the differentially methylated region upstream of the human H19 gene. J Forensic Sci 48:1275–1279PubMed Naito E, Dewa K, Fukuda M, Sumi H, Wakabayashi Y, Umetsu K, Yuasa I, Yamanouchi H (2003) Novel paternity testing by distinguishing parental alleles at a VNTR locus in the differentially methylated region upstream of the human H19 gene. J Forensic Sci 48:1275–1279PubMed
34.
go back to reference Nakayashiki N, Kanetake J, Aoki Y (2004) A parent-of-origine detectable polymorphism in the hypermethylated region upstream of the human H19 gene. Int J Leg Med 118:158–161CrossRef Nakayashiki N, Kanetake J, Aoki Y (2004) A parent-of-origine detectable polymorphism in the hypermethylated region upstream of the human H19 gene. Int J Leg Med 118:158–161CrossRef
Metadata
Title
Potential forensic application of DNA methylation profiling to body fluid identification
Authors
Hwan Young Lee
Myung Jin Park
Ajin Choi
Ja Hyun An
Woo Ick Yang
Kyoung-Jin Shin
Publication date
01-01-2012
Publisher
Springer-Verlag
Published in
International Journal of Legal Medicine / Issue 1/2012
Print ISSN: 0937-9827
Electronic ISSN: 1437-1596
DOI
https://doi.org/10.1007/s00414-011-0569-2

Other articles of this Issue 1/2012

International Journal of Legal Medicine 1/2012 Go to the issue

Editorial

Editorial