CaDNAP: Canine DNA Profiling in forensic casework

If you think about DNA analysis for CSI (crime scene investigation) the involved humans (victim, suspect) certainly come first place. Their DNA profiles can be used to potentially prove their presence at the crime scene and their relation to the crime. Also, non-human DNA may be of equally important evidentiary value and the chance of finding relevant animal DNA is quite high given the strong social interaction of humans with pets. A successful screening method to distinguish human from non-human DNA was established earlier by analysis of the cytochrom b (cytb) gene [Parson 2000] and in some cases the determination of the biological species suffices to give the relevant answers. Sometimes however, the discrimination of individuals within a species becomes important, such as is the case with popular animals. In that sense the dog can be considered the most interesting animal species from a forensic view point. The dog is deemed to be our closest animal companion and most popular pet demonstrated by more than 400 dog breeds that are known to share people’s homes. As a consequence of the high abundance and the close integration of dogs into human social life, forensically relevant cases involving dogs, such as accidents or dog attacks, are observed regularly. Even more importantly canine tissues can serve as evidentiary link when they indicate the suspect's (or victim's) presence at the crime scene. We have been carrying out research on canine DNA profiling since 2001 by introducing molecular technology to aid the canine identification process [Eichmann 2004, 2005, 2006, Hellmann 2006, Berger 2014].

Canine STRs: Allelic ladders

Allelic ladders

Example of a Canine STR- Multiplex

Canine STR-Multiplex

Canine mtDNA CR

mtDNA controlregion

mtDNA controlregion sequence

Canine DNA Profiling (CaDNAP) group

To drive international harmonization of analysis and data interpretation we co-founded the Canine DNA Profiling (CaDNAP) group in 2003 in collaboration with the German Federal Criminal Police Office (Bundeskriminalamt Wiesbaden [BKA]). In 2008 the Institute of Veterinary Pathology, Justus-Liebig-University, Giessen (Institut für Veterinärpathologie) joined our group, in 2015 the Institute of Forensic Medicine, University of Zurich (Institut für Rechtsmedizin Zürich).

The attempts of the CaDNAP group have meanwhile been recognized by other communities, such as the International Society for Animal Genetics (ISAG) who adopted some of the recommendations for canine DNA identity testing [Budowle 2005]. In 2011 members of the CaDNAP group were invited as co-authors to publish the recommendations of the International Society for Forensic Genetics regarding the use of non-human (animal) DNA in forensic genetic investigations [Linacre 2011].

In 2014 the CaDNAP group published protocols for the analysis of thirteen dog specific STRs and two sex-specific markers, assembled in two multiplex reactions that were validated according to the above mentioned ISFG guidelines [Berger 2014]. Population genetic parameters were calculated for these markers based on 295 dog samples collected in Austria (124) and Germany (171). A repeat-based nomenclature of the mainly tetrameric STRs and corresponding allelic ladders are presented in this paper. All 146 different alleles included in the ladders were sequenced for correct allele calling. Additionally, the permanent canine cell line (DH82-D3167) was evaluated as standard reference material. Alleles of this sample were designated by direct comparison to the allelic ladders and by sequence analysis of the repeat array and the flanking upstream and downstream region of all alleles. Additionally, the mitochondrial DNA haplotype of the cell line was determined by sequencing the hypervariable segments of the control region (CR) by two different laboratories independently. The difference-coded annotation was performed with respect to the generally accepted reference sequence (U96639.2).

The analysis of the canine mtDNA control region (CR) is often the last resort for DNA typing when nuclear DNA analysis fails to give useful results. This is particularly the case for the analysis of canine hairs that represent the most common crime scene evidence originating from dogs. According to the notation guidelines for human mtDNA variations of the canine mtDNA are recorded by the differences to a reference sequence (U96639.2 or NC_002008).

CaDNAP Group
U. Rohleder [BKA], U. Schleenbecker [BKA], N. Morf [RMZ], W. Hecht [Giessen], N. Schury [BKA], C. Berger [GMI], B. Berger [GMI], W. Parson [GMI]

Dr. Andreas Hellmann, Dr. Uwe Schleenbecker, Udo Rohleder
Bundeskriminalamt Wiesbaden,  Kriminaltechnisches Institut
KT 32 - Pflanzen-, Tier- und Bodenspuren

Dr. Werner Hecht
Institut für Veterinärpathologie, Giessen

Nadja Morf, MSc
Institut für Rechtsmedizin Zürich

Proficiency tests

Proficiency tests including STR and mtDNA analyses are organized every second year. A description of the proficiency test 2014/2015 was presented as a poster at the ISFG meeting in Krakow 2015.

CaDNAP Proficiency Test 2016/2017

The CaDNAP Proficiency Test 2016/2017 is now open for registration. Time for applications ends on 20th December, 2016.

CaDNAP Meetings

The group has organized meetings to harmonize scientific work and developments in the canine forensic genetic field:

First Meeting „Forensic canine DNA profilingWiesbaden 14.-15.05.2003

Second Meeting „Forensic canine DNA profilingGöttingen 23.-24.07.2003

Third Meeting: „Forensic canine DNA profilingInnsbruck 16.-18.02.2004

Fourth Meeting: „Forensic canine DNA profilingWiesbaden 03.-05.11.2004

Fifth Meeting: „Forensic canine DNA profilingKassel 01.-02.06.2006

Sixth Meeting: „Forensic canine DNA profilingInnsbruck 30.06.-02.07.2008

Seventh Meeting: „Forensic canine DNA profiling“ Giessen 30.06.-01.07.2009

Eighth Meeting: „Forensic canine DNA profiling“ Wiesbaden 29.06.-30.06.2010

Ninth Meeting: „Forensic canine DNA profiling“ Innsbruck 08.07.-10.07.2013

Tenth Meeting: „Forensic canine DNA profiling“ Giessen 15.07.-17.07.2014

Eleventh Meeting: „Forensic canine DNA profiling“ Innsbruck 28.09.-30.09.2015

Twelfth Meeting: „Forensic canine DNA profiling“ Zürich 18.07.-20.07.2016

Literature Cited

Berger B 2014 FSI Genetics 8(1): 90
Berger C 2009 Forensic Sci Rev 21:1
Budowle B 2005 Int J Legal Med 119(5): 395
Eichmann C 2004 Int J Legal Med 118(5): 249
Eichmann C 2004 Int J Legal Med 118(6): 337
Eichmann C 2005 Forensic Sci Int 151(1): 37
Eichmann C 2007 Int J Legal Med 121(5): 411
Hellmann A 2006 J Forensic Sci 51(2): 274
Linacre A 2011 FSI Genetics 5(5): 501
Parson W 2000 Int J Legal Med 114 (1-2): 23

Bibliography

Linacre,A.; Gusmao,L.; Hecht,W.; Hellmann,A.P.; Mayr,W.R.; Parson,W.; Prinz,M.; Schneider,P.M.; Morling,N.;
Budowle,B.; Garofano,P.; Hellman,A.; Ketchum,M.; Kanthaswamy,S.; Parson,W.; van Haeringen,W.; Fain,S.; Broad,T.;
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