Abstract
The differentiation of blood and menstrual fluid is especially important in cases of alleged sexual assault. While the identification of blood is relatively straightforward, the identification of menstrual fluid in trace evidence has been shown to be more challenging. This may be due to the complex nature of menstrual fluid that leads to intra- and inter-individual differences in composition. Nevertheless, recent advances in DNA methylation profiling have revealed promising markers for the differentiation of the two body fluids and furthermore, markers to distinguish menstrual fluid from vaginal fluid. A literature study was performed and in total, 11 markers were evaluated in this study of which seven could be validated for menstrual fluid and blood identification purposes. Marker “BLU2” (chr16:29757334) was identified as most suitable for differentiation of blood and menstrual fluid.
Similar content being viewed by others
References
van den Berge M, Carracedo A, Gomes I, Graham EAM, Haas C, Hjort B, Hoff-Olsen P, Maroñas O, Mevåg B, Morling N, Niederstätter H, Parson W, Schneider PM, Court DS, Vidaki A, Sijen T (2014) A collaborative European exercise on mRNA-based body fluid/skin typing and interpretation of {DNA} and {RNA} results. Forensic Sci. Int. Genet. 10:40–48. https://doi.org/10.1016/j.fsigen.2014.01.006
Holtkötter H, Beyer V, Schwender K, Glaub A, Johann KS, Schürenkamp M, Sibbing U, Banken S, Wiegand P, Pfeiffer H, Dennany L, Vennemann M (2017) EUROFORGEN-NoE Consortium, Independent validation of body fluid-specific CpG markers and construction of a robust multiplex assay. Forensic Sci Int Genet 29:261–268. https://doi.org/10.1016/j.fsigen.2017.05.002.
Olive D, Palter S (n.d.) Reproductive physiology, 14th edn. Berek and Novak’s gynecology, Philadelphia
Sensabaugh G (n.d.) Encyclopedia of forensic and legal medicine, Academic Press
Baker DJ, Grimes EA, Hopwood AJ (2011) D-dimer assays for the identification of menstrual blood. Forensic Sci Int 212(1-3):210–214. https://doi.org/10.1016/j.forsciint.2011.06.013
Holtkötter H, Dierig L, Schürenkamp M, Sibbing U, Pfeiffer H, Vennemann M (2015) Validation of an immunochromatographic D-dimer test to presumptively identify menstrual fluid in forensic exhibits. Int J Legal Med 129(1):37–41. https://doi.org/10.1007/s00414-014-1097-7
Roeder AD, Haas C (2013) mRNA profiling using a minimum of five mRNA markers per body fluid and a novel scoring method for body fluid identification. Int J Legal Med 127(4):707–721. https://doi.org/10.1007/s00414-012-0794-3
Haas C, Hanson E, Anjos MJ, Ballantyne KN, Banemann R, Bhoelai B, Borges E, Carvalho M, Courts C, De Cock G, Drobnic K, Dötsch M, Fleming R, Franchi C, Gomes I, Hadzic G, Harbison SA, Harteveld J, Hjort B, Hollard C, Hoff-Olsen P, Hüls C, Keyser C, Maroñas O, McCallum N, Moore D, Morling N, Niederstätter H, Noël F, Parson W, Phillips C, Popielarz C, Roeder AD, Salvaderi L, Sauer E, Schneider PM, Shanthan G, Court DS, Turanská M, van Oorschot RAH, Vennemann M, Vidaki A, Zatkalíková L, Ballantyne J (2014) RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains: results of a fourth and fifth collaborative EDNAP exercise. Forensic Sci. Int. Genet. 8(1):203–212. https://doi.org/10.1016/j.fsigen.2013.09.009
Forat S, Huettel B, Reinhardt R, Fimmers R, Haidl G, Denschlag D, Olek K (2016) Methylation markers for the identification of body fluids and tissues from forensic trace evidence. PLoS One 11(2):e0147973. https://doi.org/10.1371/journal.pone.0147973
Vidaki A, Giangasparo F, Syndercombe Court D (2016) Discovery of potential DNA methylation markers for forensic tissue identification using bisulphite pyrosequencing. Electrophoresis 37(21):2767–2779. https://doi.org/10.1002/elps.201600261
Herman JG, Graff JR, Myöhänen S, Nelkin BD, Baylin SB, Methylation-specific PCR (1996) A novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci 93:9821–9826. http://www.pnas.org/content/93/18/9821.abstract. https://doi.org/10.1073/pnas.93.18.9821
Lee HY, An JH, Jung S-E, Oh YN, Lee EY, Choi A, Yang WI, Shin K-J (2015) Genome-wide methylation profiling and a multiplex construction for the identification of body fluids using epigenetic markers. Forensic Sci Int Genet 17:17–24. https://doi.org/10.1016/j.fsigen.2015.03.002.
Lee HY, Jung S-E, Lee EH, Yang WI, Shin K-J (2016) DNA methylation profiling for a confirmatory test for blood, saliva, semen, vaginal fluid and menstrual blood, Forensic Sci. Int. Genet 24:75–82. https://doi.org/10.1016/j.fsigen.2016.06.007
Vidaki A, Johansson C, Giangasparo F (2017) Differentially methylated embryonal Fyn-associated substrate (EFS) gene as a blood-specific epigenetic marker and its potential application in forensic casework. Forensic Sci Int Genet 29:165–173. https://doi.org/10.1016/j.fsigen.2017.04.010
Lee HY, Park MJ, Choi A, An JH, Yang WI, Shin K-J (2012) Potential forensic application of DNA methylation profiling to body fluid identification. Int J Legal Med 126(1):55–62. https://doi.org/10.1007/s00414-011-0569-2
Park J-L, Kwon O-H, Kim JH, Yoo H-S, Lee H-C, Woo K-M, Kim S-Y, Lee S-H, Kim YS (2014) Identification of body fluid-specific DNA methylation markers for use in forensic science. Forensic Sci Int Genet 13:147–153. https://doi.org/10.1016/j.fsigen.2014.07.011
Kaminsky Z, Petronis A ( 2009) Methylation SNaPshot: a method for the quantification of site-specific DNA methylation levels BT-DNA methylation: methods and protocols, In: Tost J (Ed.), Humana Press, Totowa, pp. 241–255. https://doi.org/10.1007/978-1-59745-522-0_18.
Durnell Schuiling K, Likis FE (2013) Women’s gynecologic health, 2nd ed., Jones and Bartlett Publishers, Inc
Hoffman B, Schorge J, Schaffer J, Halvorson L, Bradshaw K, Cunningham F (2012) Williams Gynecology, in: Second Ed., McGraw Hill Professionals
Wathne B, Holst E, Hovelius B, Mårdh P-A (1994) Vaginal discharge - comparison of clinical, laboratory and microbiological findings. Acta Obstet Gynecol Scand 73(10):802–808. https://doi.org/10.3109/00016349409072509
Acknowledgements
We thank all donors for their participation in this project and the research group at King’s College London for providing us with additional semen samples. We also thank Kristina Schulze Johann, Sabrina Banken, Marianne Schürenkamp, and Ulla Sibbing for supporting this project.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Table S1
(PDF 503 kb)
Rights and permissions
About this article
Cite this article
Holtkötter, H., Schwender, K., Wiegand, P. et al. Marker evaluation for differentiation of blood and menstrual fluid by methylation-sensitive SNaPshot analysis. Int J Legal Med 132, 387–395 (2018). https://doi.org/10.1007/s00414-018-1770-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00414-018-1770-3