Highlights
- •EDNAP organized a collaborative exercise on RNA/DNA co-analysis of skin samples.
- •8 skin and 3 HKG markers were tested for specificity, sensitivity and applicability.
- •LCE1C, LOR and B2M were detected in most contact traces, detection of the other markers was inconsistent.
- •LCE1C and LOR markers showed some cross-reactivity with non-skin sample types.
- •With contact traces the method reaches the detection limit.
Abstract
The European DNA profiling group (EDNAP) organized a sixth collaborative exercise
on RNA/DNA co-analysis for body fluid/tissue identification and STR profiling. The
task was to identify skin samples/contact traces using specific RNA biomarkers and
test three housekeeping genes for their suitability as reference genes. Eight stains,
a skin RNA dilution series and, optionally, bona fide or mock casework samples of
human or non-human origin were analyzed by 22 participating laboratories using RNA
extraction or RNA/DNA co-extraction methods. Two sets of previously described skin-specific
markers were used: skin1 pentaplex (LCE1C, LCE1D, LCE2D, IL1F7 and CCL27) and skin2
triplex (LOR, KRT9 and CDSN) in conjunction with a housekeeping gene, HKG, triplex
(B2M, UBC and UCE). The laboratories used different chemistries and instrumentation.
All laboratories were able to successfully isolate and detect mRNA in contact traces
(e.g., human skin, palm-, hand- and fingerprints, clothing, car interiors, computer
accessories and electronic devices). The simultaneous extraction of RNA and DNA provides
an opportunity for positive identification of the tissue source of origin by mRNA
profiling as well as a simultaneous identification of the body fluid donor by STR
profiling. The skin markers LCE1C and LOR and the housekeeping gene marker B2M were
detected in the majority of contact traces. Detection of the other markers was inconsistent,
possibly due to the low amounts and/or poor quality of the genetic material present
in shed skin cells. The results of this and the previous collaborative RNA exercises
support RNA profiling as a reliable body fluid/tissue identification method that can
easily be combined with current STR typing technology.
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Forensic Science International: GeneticsAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- A novel histological technique for distinguishing between epithelial cells in forensic casework.Forensic Sci. Int. 2008; 178: 1-6
- The use of Polilight in the detection of seminal fluid, saliva, and bloodstains and comparison with conventional chemical-based screening tests.J. Forensic Sci. 2006; 51: 361-370
- Detection of epithelial cells in dried blood stains by reverse transcriptase-polymerase chain reaction.J. Forensic Sci. 1999; 44: 1232-1236
- Evaluation of mRNA markers for the identification of menstrual blood.J. Forensic Sci. 2002; 47: 1278-1282
- A method for simultaneous RNA and DNA isolation from dried blood and semen stains.Forensic Sci. Int. 2003; 136: 76-78
- Protamine mRNA as molecular marker for spermatozoa in semen stains.Int. J. Legal Med. 2003; 117: 175-179
- Quantification of RNA degradation by semi-quantitative duplex and competitive RT-PCR: a possible indicator of the age of bloodstains?.Forensic Sci. Int. 2003; 138: 94-103
- Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification.Forensic Sci. Int. 2003; 135: 85-96
- An mRNA and DNA co-isolation method for forensic casework samples.Anal. Biochem. 2004; 335: 289-298
- Multiplex mRNA profiling for the identification of body fluids.Forensic Sci. Int. 2005; 152: 1-12
- Messenger RNA profiling: a novel method for body fluid identification by real-time PCR.Forensic Sci. Int. 2006; 157: 181-186
- Validity of messenger RNA expression analyses of human saliva.Clin. Cancer Res. 2007; 13: 1351
- RNA in forensic science.Forensic Sci. Int. Genet. 2007; 1: 69-74
- mRNA profiling for body fluid identification by multiplex quantitative RT-PCR.J. Forensic Sci. 2007; 52: 1252-1262
- Identification of menstrual blood by real time RT-PCR: technical improvements and the practical value of negative test results.Forensic Sci. Int. 2008; 174: 55-59
- Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains.J. Forensic Sci. 2008; 53: 296-305
- 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
- mRNA profiling for body fluid identification by reverse transcription endpoint PCR and realtime PCR.Forensic Sci. Int. Genet. 2009; 3: 80-88
- mRNA profiling for the identification of sperm and seminal plasma.Forensic Sci. Int. Genet. Suppl. Ser. 2009; 2: 534-535
- How specific are the vaginal secretion markers HBD1 and MUC4?.Forensic Sci. Int. Genet. Suppl. Ser. 2009; 2: 536-537
- Evaluation of mRNA-based approach for identification of saliva and semen.Legal Med. (Tokyo). 2009; 11: 125-128
- 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
- The use of bacteria for the identification of vaginal secretions.Forensic Sci. Int. Genet. 2010; 4: 311-315
- The development of a mRNA multiplex RT-PCR assay for the definitive identification of body fluids.Forensic Sci. Int. Genet. 2010; 4: 244-256
- Detection of dermcidin for sweat identification by real-time RT-PCR and ELISA.Forensic Sci. Int. 2010; 194: 80-84
- Selection of highly specific and sensitive mRNA biomarkers for the identification of blood.Forensic Sci. Int. Genet. 2011; 5: 449-458
- Expression of statherin mRNA and protein in nasal and vaginal secretions.Legal Med. (Tokyo). 2011; 13: 309-313
- Trials of the detection of semen and vaginal fluid RNA using the genome profiling method.Legal Med. (Tokyo). 2011; 13: 265-267
- Detection of bacterial 16 S ribosomal RNA genes for forensic identification of vaginal fluid.Legal Med. (Tokyo). 2012; 14: 160-162
- Body fluid identification in forensics.BMB Rep. 2012; 45: 545-553
- Capillary electrophoresis of a multiplex reverse transcription-polymerase chain reaction to target messenger RNA markers for body fluid identification.Methods Mol. Biol. 2012; 830: 169-183
- A multiplex (m)RNA-profiling system for the forensic identification of body fluids and contact traces.Forensic Sci. Int. Genet. 2012; 6: 565-577
- Evaluation of mRNA marker specificity for the identification of five human body fluids by capillary electrophoresis.Forensic Sci. Int. Genet. 2012; 6: 452-460
- 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. 2012; : 707-721
- Successful mRNA profiling of 23 years old blood stains.Forensic Sci. Int. Genet. 2012; 6: 274-276
- Highly specific mRNA biomarkers for the identification of vaginal secretions in sexual assault investigations.Sci. Justice. 2013; 53: 14-22
- mRNA profiling for vaginal fluid and menstrual blood identification.Forensic Sci. Int. Genet. 2013; 7: 272-278
- DNA fingerprinting secondary transfer from different skin areas: morphological and genetic studies.Forensic Sci. Int. Genet. 2014; 11: 137-143
- Specific and sensitive mRNA biomarkers for the identification of skin in ‘touch DNA' evidence.Forensic Sci. Int. Genet. 2012; 6: 548-558
- mRNA-based skin identification for forensic applications.Int. J. Legal Med. 2011; 125: 253-263
- Large-scale analysis of the human and mouse transcriptomes.Proc. Nat. Acad. Sci. U. S. A. 2002; 99: 4465-4470
- mRNA profiling for the identification of blood-results of a collaborative EDNAP exercise.Forensic Sci. Int. Genet. 2011; 5: 21-26
- RNA/DNA co-analysis from blood stains – results of a second collaborative EDNAP exercise.Forensic Sci. Int. Genet. 2012; 6: 70-80
- RNA/DNA co-analysis from human saliva and semen stains – results of a third collaborative EDNAP exercise.Forensic Sci. Int. Genet. 2013; 7: 230-239
- 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. 2014; 8: 203-212
- Simplified low-copy-number DNA analysis by post-PCR purification.J. Forensic Sci. 2007; 52: 820-829
- On RNA profiling and RNA data interpretation – results of a collaborative EUROFORGEN exercise.Forensic Sci. Int. Genet.2014
- Can mRNA markers distinguish traces generated by different types of contact?.Forensic Sci. Int. Genet. Suppl. Ser. 2013; 4: e3-e4
Maximizing the performance of capillary electrophoresis systems, Applied Biosystems, Forensic News, July 2010, p27-33, http://www.lifetechnologies.com/content/dam/LifeTech/migration/us-media-library/product---services/services/pdf.par.90589.file.dat/forensic-news-july-2010.pdf.
- Good shedder or bad shedder – the influence of skin diseases on forensic DNA analysis from epithelial abrasions.Int. J. Legal Med. 2012; 126: 179-183
Article info
Publication history
Published online: January 07, 2015
Accepted:
January 4,
2015
Received in revised form:
December 12,
2014
Received:
September 24,
2014
Identification
Copyright
© 2015 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
