International Journal of Legal Medicine

, Volume 127, Issue 5, pp 891–900 | Cite as

Development of a mRNA profiling multiplex for the inference of organ tissues

  • Alexander Lindenbergh
  • Margreet van den Berge
  • Roelof-Jan Oostra
  • Cindy Cleypool
  • Annette Bruggink
  • Ate Kloosterman
  • Titia SijenEmail author
Original Article


Forensic characterisation of organ tissue generally occurs through histological and immunological assays of limited sensitivity. Here, we explore an alternative approach and examine a total of 41 candidate mRNA markers for their ability to differentiate between brain, lung, liver, skeletal muscle, heart, kidney and skin. Various selection rounds are applied involving 85 organ tissues (36 excised autopsy specimens and 49 frozen tissue sections, with at least ten specimens for each organ type), 20 commercially available RNAs from different human tissues and at least two specimens of blood, saliva, semen, vaginal mucosa, menstrual secretion or touch samples. Finally, 14 markers are regarded tissue-specific and included in an endpoint RT-PCR multiplex together with one general muscle, one blood and one housekeeping marker. This 17-plex is successfully used to analyse a blind test set of 20 specimens including mixtures, and samples derived from stabbing of organ tissues. With the blind test set samples, it is shown that an earlier described interpretation strategy for RNA cell typing results [1] is also effective for tissue inference. As organ-typing is embedded in a procedure of combined DNA/RNA extraction and analysis, both donor and organ type information is derived from the same sample. Some autopsy specimens presented DNA profiles characteristic for degraded DNA. Nevertheless, the organ-typing multiplex could generate full RNA profiles, which is probably due to small sizes of the amplicons. This assay provides a novel tool for analysis of samples from violent crimes.


Forensic science RNA profiling Organ typing Tissue identification Autopsy specimens 



The authors are very grateful to all donors from whom tissues have been used in this study. We thank Kees Krijgsman and Eppo van Houten for excising the autopsy samples. Role of funding: this study was supported by a grant from the Netherlands Genomics Initiative (NGI)/Netherlands Organization for Scientific Research (NWO) within the framework of the Forensic Genomics Consortium Netherlands (FGCN). The work leading to these results also received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 285487 (EUROFORGEN-NoE). We thank Lindy Clarisse for technical assistance and Antoinette Westen for assistance with the manuscript.

Supplementary material

414_2013_895_MOESM1_ESM.doc (376 kb)
ESM 1 (DOC 375 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Alexander Lindenbergh
    • 1
  • Margreet van den Berge
    • 1
  • Roelof-Jan Oostra
    • 2
  • Cindy Cleypool
    • 3
  • Annette Bruggink
    • 4
  • Ate Kloosterman
    • 1
  • Titia Sijen
    • 1
    Email author
  1. 1.Department of Human Biological TracesNetherlands Forensic InstituteThe HagueThe Netherlands
  2. 2.Department of Anatomy Embryology and Physiology, Academic Medical CenterUniversity of AmsterdamAmsterdamThe Netherlands
  3. 3.Department of AnatomyUniversity Medical Center UtrechtUtrechtThe Netherlands
  4. 4.UMC Utrecht BiobankUniversity Medical Center UtrechtUtrechtThe Netherlands

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