Abstract
Body fluid identification is a substantial part of forensic trace analyses. The correct determination of the origin of a biological stain may give valuable information regarding the circumstances of a crime. A simple way to detect a body fluid in a stain is the use of immunochromatographic strip tests. They are easy to use, user-independent, quick, and cheap. Currently, however, it is only possible to analyze one body fluid at a time, requiring the analyst to make previous, possibly subjective, assumptions on the body fluid at hand. Also, identification of mixed body fluids requires the use of several tests, which results in additional sample and time consumption. To combine a simple approach with the possibility to simultaneously detect several body fluids, we constructed a combined immunochromatographic strip test array based on commercially available tests. The array rapidly detects up to five body fluids with a single analysis, and allowing for subsequent DNA extraction from the same material. With this test it was possible to identify the components of a mixture, the test was easily incorporated into standard laboratory work, and its sensitivity and specificity were shown to be comparable to those of conventional strip tests.
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References
An JH, Shin KJ, Yang WI, Lee HY (2012) Body fluid identification in forensics. BMB Rep
Tobe SS, Watson N, Nic Daéid N (2007) Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA. J Forensic Sci 52:102–109. https://doi.org/10.1111/j.1556-4029.2006.00324.x
Holtkötter H, Dierig L, Schürenkamp M, Sibbing U, Pfeiffer H, Vennemann M (2014) Validation of an immunochromatographic D-dimer test to presumptively identify menstrual fluid in forensic exhibits. Int J Legal Med 129:37–41
Baker DJ, Grimes EA, Hopwood AJ (2011) D-dimer assays for the identification of menstrual blood. Forensic Sci Int 212:210–214. https://doi.org/10.1016/j.forsciint.2011.06.013
Sikirzhytski V, Virkler K, Lednev IK (2010) Discriminant analysis of Raman spectra for body fluid identification for forensic purposes. Sensors (Basel) 10:2869–2884. https://doi.org/10.3390/s100402869
Muro CK, Doty KC, de Souza Fernandes L, Lednev IK (2016) Forensic body fluid identification and differentiation by Raman spectroscopy. Forensic Chem 1:31–38. https://doi.org/10.1016/j.forc.2016.06.003
Zapata F, Gregorio I, Garcia-Ruiz C (2016) Body fluids and spectroscopic techniques in forensics: a perfect match? J Forensic Med 1:1–7. https://doi.org/10.4172/2472-1026.1000101
Durnell Schuiling K, Likis FE (2013) Women’s gynecologic health. 2nd ed. Jones and Bartlett Publishers, Inc
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 188:1–17. https://doi.org/10.1016/j.forsciint.2009.02.013
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 8:203–212. https://doi.org/10.1016/j.fsigen.2013.09.009
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
Juusola J, Ballantyne J (2003) Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification. Forensic Sci Int 135:85–96. https://doi.org/10.1016/S0379-0738(03)00197-X
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 157:181–186. https://doi.org/10.1016/j.forsciint.2005.10.009
Richard MLL, Harper KA, Craig RL, Onorato AJ, Robertson JM, Donfack J (2012) Evaluation of mRNA marker specificity for the identification of five human body fluids by capillary electrophoresis. Forensic Sci Int Genet 6:452–460. https://doi.org/10.1016/j.fsigen.2011.09.007
Rekker K, Saare M, Roost AM, Salumets A, Peters M (2013) Circulating microRNA profile throughout the menstrual cycle. PLoS One 8:e81166. https://doi.org/10.1371/journal.pone.0081166
Ohlsson Teague EMC, Print CG, Hull ML (2010) The role of microRNAs in endometriosis and associated reproductive conditions. Hum Reprod Update 16:142–165. https://doi.org/10.1093/humupd/dmp034
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
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:e22. https://doi.org/10.1371/journal.pbio.0060022
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:e0147973. https://doi.org/10.1371/journal.pone.0147973
An JH, Choi A, Shin K-J, Yang WI, Lee HY (2013) DNA methylation-specific multiplex assays for body fluid identification. Int J Legal Med 127:35–43. https://doi.org/10.1007/s00414-012-0719-1
Frumkin D, Wasserstrom A, Budowle B, Davidson A (2011) DNA methylation-based forensic tissue identification. Forensic Sci Int Genet 5:517–524. https://doi.org/10.1016/j.fsigen.2010.12.001
Holtkötter H, Beyer V, Schwender K, Glaub A, Johann KS, Schürenkamp M, Sibbing U, Banken S, Wiegand P, Pfeiffer H, Dennany L, Vennemann M, EUROFORGEN-NoE Consortium (2017) Independent validation of body fluid-specific CpG markers and construction of a robust multiplex assay. Forensic Sci Int Genet. https://doi.org/10.1016/j.fsigen.2017.05.002
Harbison S, Fleming R (2016) Forensic body fluid identification: state of the art. Res Reports Forensic Med Sci 6:11. https://doi.org/10.2147/RRFMS.S57994
Turrina S, Filippini G, Atzei R, Zaglia E, De Leo D (2008) Validation studies of rapid stain identification-blood (RSID-blood) kit in forensic caseworks. Forensic Sci Int Genet Suppl Ser 1:74–75. https://doi.org/10.1016/j.fsigss.2007.10.166
SERATEC®, SERATEC® PMB user instruction (2017) 1–2. http://www.seratec.com/docs/user_instructions/PMB_en.pdf
SERATEC®, SERATEC HemDirect (2013)
Old JB, Schweers BA, Boonlayangoor PW, Reich KA (2009) Developmental validation of RSID™-Saliva: a lateral flow immunochromatographic strip test for the forensic detection of saliva. J Forensic Sci 54:866–873. https://doi.org/10.1111/j.1556-4029.2009.01055.x
SERATEC®, SERATEC α-Amylase test (2013)
Independent Forensics, Developmental validation of RSID-urine, n.d. http://www.galantos.eu/downloads/RSID/Validation-urine-rsid.pdf
Old J, Schweers BA, Boonlayangoor PW, Fischer B, Miller KWP, Reich K (2012) Developmental validation of RSID™-semen: a lateral flow Immunochromatographic strip test for the forensic detection of human semen*. J Forensic Sci 57:489–499. https://doi.org/10.1111/j.1556-4029.2011.01968.x
SERATEC®, SERATEC PSA Semiquant (2013). http://www.seratec.com/docs/user_instructions/psm400f_multi.pdf
Independent Forensics (n.d.) Rapid stain identification of human saliva (RSID™-Saliva)—technical information and protocol sheet for use with universal buffer, reduced incubation time Cat# 0130. http://www.galantos.eu/downloads/RSID/102015short incubSALIVA-Universal Buffer.pdf
Independent Forensics (n.d.) Rapid stain identification of urine (RSID™-urine)—technical information and protocol sheet for cat# 0400
Independent Forensics (n.d.) Rapid stain identification of human semen (RSID™-Semen)—technical information and protocol sheet for use with universal buffer, reduced incubation time, cat# 0230, 1–2. http://www.galantos.eu/downloads/RSID/protocols-semen/101915 short incub SEMEN-Univ Buffer.pdf
Sato I, Sagi M, Ishiwari A, Nishijima H, Ito E, Mukai T (2002) Use of the “SMITEST” PSA card to identify the presence of prostate-specific antigen in semen and male urine. Forensic Sci Int 127:71–74. https://doi.org/10.1016/S0379-0738(02)00111-1
Hooft PJ, Van De Voorde HP (1990) The zinc test as an alternative for acid phosphatase spot tests in the primary identification of seminal traces. Forensic Sci Int 47:269–275. https://doi.org/10.1016/0379-0738(90)90296-B
Schmidt S, Franke M, Lehmann J, Loch T, Stöckle M, Weichert-Jacobsen K (2001) Prostate-specific antigen in female urine: a prospective study involving 217 women. Urology 57:717–720. https://doi.org/10.1016/S0090-4295(00)01093-1
Holtkötter H, Rodrigues Filho C, Schwender K, Stadler C, Vennemann M, Pacheco AC, Roca MG (2017) Forensic differentiation between peripheral and menstrual blood in cases of alleged sexual assault—validating an immunochromatographic multiplex assay for simultaneous detection of human hemoglobin and D-dimer. Int J Leg Med
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We thank all donors for their participation in this project. We also thank Kristina Schulze Johann, Sabrina Banken, Marianne Schürenkamp, and Ulla Sibbing for supporting this project.
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Body fluid samples were collected with informed consent using procedures approved by the local ethical committee (Ethik-Kommission der Ärztekammer Westfalen-Lippe und der Westfälischen Wilhelms-Universität Münster).
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Holtkötter, H., Schwender, K., Wiegand, P. et al. Improving body fluid identification in forensic trace evidence—construction of an immunochromatographic test array to rapidly detect up to five body fluids simultaneously. Int J Legal Med 132, 83–90 (2018). https://doi.org/10.1007/s00414-017-1724-1
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DOI: https://doi.org/10.1007/s00414-017-1724-1