Determination of body fluid based on analysis of nucleic acids

Authors: Marie Korabečná 1,2
Authors‘ workplace: Ústav biologie a lékařské genetiky 1. lékařské fakulty UK a Všeobecné fakultní nemocnice, Praha 1;  Lékařská fakulta UK v Plzni, Plzeň 2
Published in: Soud Lék., 60, 2015, No. 3, p. 33-36
Category: Review


Recent methodological approaches of molecular genetics allow isolation of nucleic acids (DNA and RNA) from negligible forensic samples. Analysis of these molecules may be used not only for individual identification based on DNA profiling but also for the detection of origin of the body fluid which (alone or in mixture with other body fluids) forms the examined biological trace. Such an examination can contribute to the evaluation of procedural, technical and tactical value of the trace. Molecular genetic approaches discussed in the review offer new possibilities in comparison with traditional spectrum of chemical, immunological and spectroscopic tests especially with regard to the interpretation of mixtures of biological fluids and to the confirmatory character of the tests. Approaches based on reverse transcription of tissue specific mRNA and their subsequent polymerase chain reaction (PCR) and fragmentation analysis are applicable on samples containing minimal amounts of biological material. Methods for body fluid discrimination based on examination of microRNA in samples provided so far confusing results therefore further development in this field is needed. The examination of tissue specific methylation of nucleotides in selected gene sequences seems to represent a promising enrichment of the methodological spectrum. The detection of DNA sequences of tissue related bacteria has been established and it provides satisfactory results mainly in combination with above mentioned methodological approaches.

body fluids – mRNA – microRNA – qPCR– promotermethylation – microbial DNA


1. Sikirzhytski V, Sikirzytskaya A, Lednev IK. Multidimensional Raman spectroscopic signatures as a tol for forensic identifiation of body fluids traces: a review. Appl Spectrosc 2011; 65: 1223-1232.

2. Scano O, Locci E, Noto A et al. 1H NMR metabolite fingerprinting as a new tool for body fluid identification in forensic science. Magn Reson Chem 2013; 51: 454-462.

3. Frascione N, Pinto V, Daniel B. Development of biosensor for human blood: new routes to body fluid identification. Anal Bioanal Chem 2012; 404: 23-28.

4. Frascione N, Thorogate R, Daniel B, Jickells S. Detection and identification of body fluids stains using antibody-nanoparticle conjugates. Analyst 2012; 137: 508-512.

5. Virkler K, Lednev IK. Analysis of body fluids for forensic purposes: from laboratory testing to non-destructive rapid confirmatory identification at a crime scene. Forensic Sci Int 2009; 188: 1-17.

6. AnJH, ShinKJ, YangWI, LeeHY. Body fluid identification in forensics. BMB Rep 2012; 45: 545-553.

7. Zubakov D, Hanekamp E, Kokshoorn M, van IJcken W, Kayser M. Stable RNA markers for identification of blood and saliva stains revealed from whole genome expression analysis of time-wise degraded samples. Int J Legal Med 2008; 122: 135-142.

8. Zubakov D, Kokshoorn M, Kloosterman A, Kayser M. New markers for old stains: stable mRNA markers for blood and saliva identification from up to16-year-old stains. Int J Legal Med 2009; 123: 71-74.

9. Setzer M, Juusola J, Ballantyne J. Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains. J Forensic Sci 2008; 53: 296-305.

10. Bauer M, Patzelt D. Simultaneous RNA and DNA isolation from blood and semen stains. Forensic Sci Int 2003; 136: 76-78.

11. Alvarez M, Juusola J, Ballantyne J. An mRNA and DNA co-isolation method for forensic casework samples. Anal Biochem 2004; 335: 289-298.

12. Haas C, Hanson E, Bär W, et al. mRNA profilig for the identification of blood-results of a collaborative EDNAP exercise. Forensic Sci Int Genet 2011; 5: 21-26.

13. Haas C, Hanson E, Anjos MJ, et al. RNA/DNA co-analysis from blood stains—results of a second collaborative EDNAP exercise. Forensic Sci Int Genet 2012; 6: 70-80.

14. Juusola J, Ballantyne J. Multiplex mRNA profilig for the identification of body fluids. Forensic Sci Int 2005; 152: 1-12.

15. Juusola J, Ballantyne J. mRNA profilig for body fluid identification by multiplex quantitative RT-PCR. J Forensic Sci 2007; 52: 1252-1262.

16. Haas C, Klesser B, Maake C, Bär W, Kratzer A. mRNA profilig for body fluid identification by reverse transcription end point PCR and realtime PCR. Forensic Sci Int Genet 2009; 3: 80-88.

17. Nussbaumer C, Gharehbaghi-Schnell E, Korschineck I. Messenger RNA profiling: a novel method for body fluid identification by real-time PCR. Forensic Sci Int 2006; 157: 181-186.

18. Xu Y, Xie J, Cao Y et al. Development of highly sensitive and specific mRNA multiplex system (XCYR1) for forensic human body fluids and tissues identification. PLoSOne 2014; 9(7): e100123.

19. Hanson EK, Lubenow H, Ballantyne J. Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs. Anal Biochem 2009; 387: 303-314.

20. Zubakov D, Boersma AW, Choi Y, van Kuijk PF, Wiemer EA, Kayser M. MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation. Int J Legal Med 2010; 124: 217-226.

21. Park JL, Park SM, Kwon OH, et al. Microarray screening and qRT-PCR evaluation of microRNA markers for forensic body fluid identification. Electrophoresis 2014; 35: 21-22.

22. Wang Z, LuoHB, Pan XF, Liao M, Hou YP. A model for data analysis of microRNA expression in forensic body fluid identification. Forensic Sci Int Genet 2012; 6: 419-423.

23. AnJH, Chi A, Shin KJ, Zang WI, Lee HY. DNA methylation-specific multiplex assays for body fluid identification. Int J Legal Med 2013; 127: 35-43.

24. Wasserstrom A, Frumkin D, Davidson A, Shpitzen M, Herman Y, Gafny R. Demonstration of DSI-semen-A novel DNA methylation-based forensic semen identification assay. Forensic Sci Int Genet 2013; 7:136-142.

25. Lee HY, Park MJ, Choi A, An JH, Yang WI, Shin KJ. Potential forensic application of DNA methylation profiling to body fluid identification. Int J Legal Med 2012; 126: 55-62.

26. Madi T, Balamurugan K, Bombardi R, Duncan G, McCord B. The determination of tissue-specific DNA methylation patterns in forensic biofluids using bisulfite modification and pyrosequencing. Electrophoresis 2012; 33: 1736-1745.

27. Park JL, Kwon OH, Kim JH, et al. Identification of body fluid-specific DNA methylation markers for use in forensic science. Forensic Sci Int Genet 2014; 13C: 147-153.

28. Hsu L, Power D, Upritchard J, et al. Amplification of oral streptococcal DNA from human incisors and bite marks. Curr Microbiol 2012; 65: 207–211.

29. Akutsu T, Motani H, Watanabe K, Iwase H, Sakurada K. Detection of bacterial 16S ribosomal RNA genes for forensic identification of vaginal fluid. Legal Med (Tokyo) 2012; 14: 160–162.

30. Giampaoli S, Berti A, Valeriani F, et al. Molecular identification of vaginal fluid by microbial signature. Forensic Sci Int Genet 2012; 6: 559–564.

31. Giampaoli S, Alessandrini F, Berti A, et al. Forensic interlaboratory evaluation of the ForFLUID kit for vaginal fluids identification. J Forensic Leg Med 2014; (21): 60-63.

32. Choi A, Shin KJ, Yang WI, Lee HY. Body fluid identification by integrated analysis of DNA methylation and body fluid-specific microbial DNA. Int J Legal Med 2014; 128: 33-41.

33. Fleming RI, Harbison S. The use of bakteria for identification of vaginal secretions. Forensic Sci Int Genet 2010; 4: 311-315.

Anatomical pathology Forensic medical examiner Toxicology
Forgotten password

Don‘t have an account?  Create new account

Forgotten password

Enter the email address that you registered with. We will send you instructions on how to set a new password.


Don‘t have an account?  Create new account