International Journal of Legal Medicine

, Volume 126, Issue 5, pp 815–823 | Cite as

Comparison of bacterial DNA profiles of footwear insoles and soles of feet for the forensic discrimination of footwear owners

  • Haruhisa GogaEmail author
Short Communication


It is crucial to identify the owner of unattended footwear left at a crime scene. However, retrieving enough DNA for DNA profiling from the owner’s foot skin (plantar skin) cells from inside the footwear is often unsuccessful. This is sometimes because footwear that is used on a daily basis contains an abundance of bacteria that degrade DNA. Further, numerous other factors related to the inside of the shoe, such as high humidity and temperature, can encourage bacterial growth inside the footwear and enhance DNA degradation. This project sought to determine if bacteria from inside footwear could be used for footwear trace evidence. The plantar skins and insoles of shoes of volunteers were swabbed for bacteria, and their bacterial community profiles were compared using bacterial 16S rRNA terminal restriction fragment length polymorphism analysis. Sufficient bacteria were recovered from both footwear insoles and the plantar skins of the volunteers. The profiling identified that each volunteer’s plantar skins harbored unique bacterial communities, as did the individuals’ footwear insoles. In most cases, a significant similarity in the bacterial community was identified for the matched foot/insole swabs from each volunteer, as compared with those profiles from different volunteers. These observations indicate the probability to discriminate the owner of footwear by comparing the microbial DNA fingerprint from inside footwear with that of the skin from the soles of the feet of the suspected owner. This novel strategy will offer auxiliary forensic footwear evidence for human DNA identification, although further investigations into this technique are required.


Bacterial DNA profiling Footwear insole Skin bacterial community Forensic Terminal restriction fragment length polymorphism 



The author thanks all of the volunteers who provided bacterial samples for this study. Special thanks to Dr. Minoru Nakazato (Lab Director of Forensic Science Division, Department of Criminal Investigation, Okinawa Prefectural Police HQ) for his encouragement during the early days of the study and to Dr. Takeshi Arakawa (Molecular Microbiology Group, Department of Tropical Infectious Diseases, COMB, Tropical Biosphere Research Center, University of the Ryukyus) for his helpful discussions on the work in this paper.


  1. 1.
    Bright J, Petricevic SF (2004) Recovery of trace DNA and its application to DNA profiling of shoe insoles. Forensic Sci Int 145:7–12PubMedCrossRefGoogle Scholar
  2. 2.
    Hillier E, Dixon P, Stewart P, Yamashita B, Lama D (2005) Recovery of DNA from shoes. Can Soc Forensic Sci J 38:143–150Google Scholar
  3. 3.
    Daly DJ, Murphy C, McDermott SD (2012) The transfer of touch DNA from hands to glass, fabric and wood. Forensic Sci Int Genet 6:41–46PubMedCrossRefGoogle Scholar
  4. 4.
    Poinar HN (2003) The top 10 list: criteria of authenticity for DNA from ancient and forensic samples. Int Congr Ser 1239:575–579CrossRefGoogle Scholar
  5. 5.
    Iwasaki F (2002) The research on the bacterial filth of the inside shoes (in Japanese with English title). J Jpn Soc Med Study Footwear 16:51–54Google Scholar
  6. 6.
    Matsuura Y (1998) The research on the effectiveness of antibacterial shoes (in Japanese). J Jpn Soc Med Study Footwear 12:27–30Google Scholar
  7. 7.
    Li H, Zhao C, Zhou J, Shao H, Chen W (2011) Isolation, purification and identification of bacteria from the shoes worn by children. Afr J Biotechnol 10:4133–4137Google Scholar
  8. 8.
    Mitsui M (2009) Effects of shoe microclimate on wear comfort (in Japanese with English title). Sen’i Gakkaishi 65:166–170Google Scholar
  9. 9.
    Butler JM, Shen Y, McCord BR (2003) The development of reduced size STR amplicons as tools for analysis of degraded DNA. J Forensic Sci 48:1054–1064PubMedGoogle Scholar
  10. 10.
    Smith PJ, Ballantyne J (2007) Simplified low-copy-number DNA analysis by post-PCR purification. J Forensic Sci 52:820–829PubMedCrossRefGoogle Scholar
  11. 11.
    Horswell J, Cordiner SJ, Maas EW, Martin TM, Sutherland KBW, Speir TW, Nogales B, Osborn AM (2002) Forensic comparison of soils by bacterial community DNA profiling. J Forensic Sci 47:350–353PubMedGoogle Scholar
  12. 12.
    Grice EA, Kong HH, Conlan S, Deming CB, Davis J, Young AC et al (2009) Topographical and temporal diversity of the human skin microbiome. Science 324:1190–1192PubMedCrossRefGoogle Scholar
  13. 13.
    Costello EK, Lauber CL, Hamady M, Fierer N, Gordon JI, Knight R (2009) Bacterial community variation in human body habitats across space and time. Science 326:1694–1697PubMedCrossRefGoogle Scholar
  14. 14.
    Fierer N, Hamady M, Lauber CL, Knight R (2008) The influence of sex, handedness, and washing on the diversity of hand surface bacteria. Proc Natl Acad Sci U S A 105:17994–17999PubMedCrossRefGoogle Scholar
  15. 15.
    Fierer N, Lauber CL, Zhou N, McDonald D, Costello EK, Knight R (2010) Forensic identification using skin bacterial communities. Proc Natl Acad Sci U S A 107:6477–6481PubMedCrossRefGoogle Scholar
  16. 16.
    Tims S, van Wamel W, Endtz HP, van Belkum A, Kayser M (2010) Microbial DNA fingerprinting of human fingerprints: dynamic colonization of fingertip microflora challenges human host inferences for forensic purposes. Int J Leg Med 124:477–481CrossRefGoogle Scholar
  17. 17.
    Liu WT, Marsh TL, Cheng H, Forney LJ (1997) Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl Environ Microbiol 63:4516–4522PubMedGoogle Scholar
  18. 18.
    Osborn AM, Moore ERB, Timmis KN (2000) An evaluation of terminal-restriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics. Environ Microbiol 2:39–50PubMedCrossRefGoogle Scholar
  19. 19.
    Dunbar J, Ticknor LO, Kuske CR (2000) Assessment of microbial diversity in four southwestern United States soils by 16S rRNA gene terminal restriction fragment analysis. Appl Environ Microbiol 66:2943–2950PubMedCrossRefGoogle Scholar
  20. 20.
    Meyers MS, Foran DR (2008) Spatial and temporal influences on bacterial profiling of forensic soil samples. J Forensic Sci 53:652–660PubMedCrossRefGoogle Scholar
  21. 21.
    Heath LE, Saunders VA (2006) Assessing the potential of bacterial DNA profiling for forensic soil comparisons. J Forensic Sci 51:1062–1068PubMedCrossRefGoogle Scholar
  22. 22.
    Knudsen EA (1986) Isolation of dermatophytes from footwear with adhesive tape strips. J Med Vet Mycol 25:59–61CrossRefGoogle Scholar
  23. 23.
    Watanabe K, Taniguchi H, Nishioka K, Maruyama R, Katoh T (2000) Preventive effects of various socks against adhesion of dermatophytes to healthy feet (in Japanese with English abstract). Nihon Ishinkin Gakkai Zasshi 41:183–186PubMedCrossRefGoogle Scholar
  24. 24.
    Maruyama R, Fukuyama K, Katoh T, Sugimoto R, Taniguchi H, Watanabe K, Nishioka K (2003) Prevention of dermatophytoses (in Japanese with English abstract). Nihon Ishinkin Gakkai Zasshi 44:265–268PubMedCrossRefGoogle Scholar
  25. 25.
    Tanaka K, Katoh T, Irimajiri J, Taniguchi H, Yokozeki H (2006) Preventive effects of various types of footwear and cleaning methods on dermatophyte adhesion. J Dermatol 33:528–536PubMedCrossRefGoogle Scholar
  26. 26.
    Abraham JH, Gold DR, Dockery DW, Ryan L, Park J, Milton DK (2005) Within-home versus between-home variability of house dust endotoxin in a birth cohort. Environ Health Perspect 113:1516–1521PubMedCrossRefGoogle Scholar
  27. 27.
    Tringe SG, Zhang T, Liu X, Yu Y, Lee WH, Yap J et al (2008) The airborne metagenome in an indoor urban environment. PLoS ONE. doi: 10.1371/journal.pone.0001862
  28. 28.
    Rintala H, Pitkäranta M, Toivola M, Paulin L, Nevalainen A (2008) Diversity and seasonal dynamics of bacterial community in indoor environment. BMC Microbiol. doi: 10.1186/1471-2180-8-56
  29. 29.
    Flores GE, Bates ST, Knights D, Lauber CL, Stombaugh J, Knight R, Fierer N (2011) Microbial biogeography of public restroom surfaces. PLoS ONE. doi: 10.1371/journal.pone.0028132
  30. 30.
    Katoh T (2006) Dermatomycosis and environment (in Japanese with English abstract). Nihon Ishinkin Gakkai Zasshi 47:63–67PubMedCrossRefGoogle Scholar
  31. 31.
    Lenz EJ, Foran DR (2010) Bacterial profiling of soil using genus-specific markers and multidimensional scaling. J Forensic Sci 55:1437–1442PubMedCrossRefGoogle Scholar
  32. 32.
    Turnbaugh PJ, Hamady M, Yatsunenko T, Cantarel BL, Duncan A, Ley RE et al (2009) A core gut microbiome in obese and lean twins. Nature 457:480–484PubMedCrossRefGoogle Scholar
  33. 33.
    Turnbaugh PJ, Quince C, Faith JJ, McHardy AC, Yatsunenko T, Niazi F et al (2010) Organismal, genetic, and transcriptional variation in the deeply sequenced gut microbiomes of identical twins. Proc Natl Acad Sci USA 107:7503–7508PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Forensic Science Division, Department of Criminal InvestigationOkinawa Prefectural Police HQNahaJapan

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