Evaluation of the suitability of mitochondrial DNA for species identification of microtraces and forensic traces.

  • Małgorzata Natonek-Wiśniewska Instytut Zootechniki PIB Dział Genomiki i Biologii Molekularnej Zwierząt
  • Piotr Krzyścin Instytut Zootechniki PIB Dział Genomiki i Biologii Molekularnej Zwierząt
Keywords: biological traces, forensic DNA analysis, species identification of forensic DNA, species identification of biological traces, mtDNA


The objective of the study was to demonstrate how mitochondrial DNA (mtDNA) can be used to determine the species origin of animal microtraces. The study included pieces of cat and dog hair without the root, a fragment of cooked chicken bone (4 ´ 4 ´ 1 mm), goose down (0.028 g), a pork swab, a pork scratching (5 ´ 5 ´ 5 mm), and pork lard (0.22 g). DNA was isolated from all of these samples using the method appropriate for a particular template. The DNA extracts had a concentration exceeding 5.4 ng/µl with A260/280 purity range of 1.14 –1.88. Next the samples were subjected to PCR and real-time PCR with species-specific primers and primers complementary to mitochondrial DNA (mtDNA). Controls based on the amplification of eukaryotic-specific fragment (18S rRNA) were additionally performed. PCR and real-time PCR products for determining species-specific mtDNA were obtained for all templates, whereas in eukaryote determination no product was obtained for dog and cat hair only.

The poor quality of the DNA obtained did not prevent the analysis. The results show that mitochondrial DNA is suitable for determination of small or highly processed samples, in which genomic DNA often cannot be analysed


Aida A, Man Y, Wong C, Raha A, Son R. (2005) Analysis of raw meats and fats of pigs using polymerase chain reaction for Halal authentication. Meat Sci. 69(1): 47-52. doi.org/10.1016/j.meatsci.2004.06.020

Bello N, Francino O, Sánchez A. (2001) Isolation of genomic DNA from feathers. J Vet Diagn Invest. 13(2): 162-164.

Di Pinto A, Forte V, Guastadisegn, M, Martino, Schena F, Tantillo G.(2007) A comparison of DNA extraction methods for food analysis. Food control 18(1): 76-80. doi.org/10.1016/j.foodcont.2005.08.011

Heywood D, Skinner R, Cornwell P (2003) Analysis of DNA in hair fibers. J Cosmetic Sci. 54(1): 21-28.

Hou B, Meng X, Zhang L, Guo J, Li S, Jin H (2015). Development of a sensitive and specific multiplex PCR method for the simultaneous detection of chicken, duck and goose DNA in meat products. Meat Sci. 101 (2015): 90-94. doi.org/10.1016/j.meatsci.2014.11.007

Hou B, Meng X, Zhang L, Guo J, Li S, Jin H. (2015). Development of a sensitive and specific multiplex PCR method for the simultaneous detection of chicken, duck and goose DNA in meat products. Meat Sci. 101: 90-94. doi.org/10.1016/j.meatsci.2014.11.007

Iniesta V, Belinchón-Lorenzo S, Soto M, Fernández-Cotrina J, Muñoz-Madrid R, Monroy I, Baz V, Gómez-Luque A, Parejo J, Alonso C, Gómez-Nieto L. (2013) Detection and chronology of parasitic kinetoplast DNA presence in hair of experimental Leishmania major infected BALB/c mice by Real Time PCR. Acta Trop. 128: 468-472. doi.org/10.1016/j.actatropica.2013.07.007

Mafra I, Silva S, Moreira E, da Silva C, Beatriz M, Oliveira P. (2008) Comparative study of DNA extraction methods for soybean derived food products. Food Control 19(12): 1183-1190. doi.org/10.1016/j.foodcont.2008.01.004

Muñoz-Madrid R, Belinchón-Lorenzo S, Iniesta V, Fernández-Cotrina J, Parejo JC, Serrano FJ, Monroy I, Ba V, Gómez-Luque A, Gómez-Nieto LC (2013). First detection of Leishmania infantum kinetoplast DNA in hair of wild mammals: application of qPCR method to determine potential parasite reservoirs. Acta Trop., 128: 706-709. doi.org/10.1016/j.actatropica.2013.08.009

Natonek-Wisniewska M, Krzyścin P (2012). Modyfikacja metody identyfikacji gatunkowej psów, bydła, świń, kur i owiec wykorzystywanej do analizy niestandardowych próbek. Roczniki Naukowe Zootechniki 39(1):17-24. http://www.izoo.krakow.pl/czasopisma/roczniki/2012/1/art02.pdf

Natonek-Wiśniewska M (2009). Species identification of feline DNA based on analysis of cytochrome b. Ann Anim Sci. 9(4): 379-383. www.izoo.krakow.pl/czasopisma/annals/2009/4/6.pdf

Natonek-Wiśniewska M, Krzyścin P, Piestrzyńska-Kajtoch A (2013) The species identification of bovine, porcine, ovine and chicken components in animal meals, feeds and their ingredients, based on COX I analysis and ribosomal DNA sequences. Food Control 34(1): 69-78. doi.org/10.1016/j.foodcont.2013.04.014

Natonek-Wiśniewska M, Krzyścin P. (2015) Opracowanie prostych i skutecznych testów real-time PCR od identyfikacji komponentów bydlęcych, wieprzowych i owczych w żywności. Zywn-Nauk Technol Ja. 22(4):73-84. DOI: 10.15193/ZNTJ/2015/101/057

Pegels, N, González I., López-Calleja I, Fernández S, García T, Martín R (2012) Evaluation of a TaqMan real-time PCR assay for detection of chicken, turkey, duck, and goose material in highly processed industrial feed samples. Poultry Sci., 91(7): 1709-1719. doi.org/10.3382/ps.2011-01954

Pfeiffer I, Völkel I, Täubert H, Brenig B (2004). Forensic DNA-typing of dog hair: DNA-extraction and PCR amplification. Forensic Sci Int. 141(2): 149-151. doi.org/10.1016/j.forsciint.2004.01.016

Vietina M, Agrimonti C, Marmiroli N. (2013). Detection of plant oil DNA using high resolution melting (HRM) post PCR analysis: A tool for disclosure of olive oil adulteration." Food chem. 141(4): 3820-3826. doi.org/10.1016/j.foodchem.2013.06.075

Yannic G, Sermier R, Aebischer A, Gavrilo M, Gilg O, Miljeteig C, Broquet T. (2011). Description of microsatellite markers and genotyping performances using feathers and buccal swabs for the Ivory gull (Pagophila eburnea). Mol. Ecol. Resour. 11:877–889. DOI: 10.1111/j.1755-0998.2011.03015