title: Development of a mRNA profiling multiplex for the inference of organ tissues.
abstract: 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.
authors.affiliation: Department of Human Biological Traces, Netherlands Forensic Institute, P.O. Box 24044, 2490 AA The Hague, The Netherlands. a.lindenbergh@nfi.minvenj.nl
authors.forename: Alexander
authors.initials: A
authors.lastname: Lindenbergh
authors.forename: Margreet
authors.initials: M
authors.lastname: van den Berge
authors.forename: Roelof-Jan
authors.initials: RJ
authors.lastname: Oostra
authors.forename: Cindy
authors.initials: C
authors.lastname: Cleypool
authors.forename: Annette
authors.initials: A
authors.lastname: Bruggink
authors.forename: Ate
authors.initials: A
authors.lastname: Kloosterman
authors.forename: Titia
authors.initials: T
authors.lastname: Sijen
mesh_terms: D001774:Blood Chemical Analysis; D001921:Brain / Q000473:pathology; D001923:Brain Chemistry; D002582:Cervix Mucus / Q000737:chemistry; D016172:DNA Fingerprinting; D017931:DNA Primers; D005260:Female; D049429:Forensic Pathology; D020869:Gene Expression Profiling; D005819:Genetic Markers; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000473:pathology; D008099:Liver / Q000737:chemistry / Q000473:pathology; D008168:Lung / Q000737:chemistry / Q000473:pathology; D008297:Male; D008598:Menstruation; D060885:Multiplex Polymerase Chain Reaction; D018482:Muscle, Skeletal / Q000737:chemistry / Q000473:pathology; D009206:Myocardium / Q000737:chemistry / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva / Q000737:chemistry; D012661:Semen / Q000737:chemistry; D012867:Skin / Q000737:chemistry / Q000473:pathology
chemical_list: D017931:DNA Primers; D005819:Genetic Markers; D012333:RNA, Messenger