International Journal of Legal Medicine

, Volume 124, Issue 6, pp 569–576 | Cite as

Using oral microbial DNA analysis to identify expirated bloodspatter

  • Andrea E. Donaldson
  • Michael C. Taylor
  • Stephen J. Cordiner
  • Iain L. Lamont


Distinguishing expirated bloodstains (blood forced by airflow out of the nose, mouth or a chest wound) from impact spatter (blood from gunshots, explosives, blunt force trauma and/or machinery accidents) is an important challenge in forensic science. Streptococcal bacteria are only found in the human mouth and saliva. This study developed a polymerase chain reaction (PCR) method that detects DNA from these bacteria as a sensitive tool to detect the presence of saliva. The PCR method was very specific to human oral streptococci, with no PCR product being made from human DNA or DNA from other microbes that were tested. It was also very sensitive, detecting as little as 60 fg of target DNA. The PCR amplification gave product with 99 out of 100 saliva samples tested. PCR was not inhibited by the presence of blood and could detect target DNA in expirated bloodstains in a range of materials and for up to 92 days after deposit on cardboard or cotton fabric. In a blind trial, the PCR method was able to distinguish three mock forensic samples that contained expirated blood from four that did not. Our data show that bacteria present in the oral cavity can be detected in bloodstains that contain saliva and therefore can potentially be used as a marker in forensic work to distinguish mouth-expirated bloodstains from other types of bloodstains.


Expirated bloodstains Saliva DNA analysis Glucosyltransferase genes Streptococcus 



We gratefully acknowledge all the willing participants who provided saliva for this research, Professor John Tagg and Megan Inglis who provided Streptococcus strains and advice on their growth and manipulation, Dr. Dan Power and Dr. Jacqui Horswell for all their suggestions and assistance with developing the project, Rachel Parkinson who provided the soil microbial DNA samples and all the other Institute of Environmental Science and Research (ESR) staff who freely gave their time, knowledge and assistance to help with this research. AED was supported by a Postgraduate Scholarship from ESR.

Supplementary material

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Supplementary material 1 (PDF 2296 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Andrea E. Donaldson
    • 1
  • Michael C. Taylor
    • 2
  • Stephen J. Cordiner
    • 3
  • Iain L. Lamont
    • 1
  1. 1.Department of BiochemistryUniversity of OtagoDunedinNew Zealand
  2. 2.Institute of Environmental Science and Research (ESR) LtdChristchurchNew Zealand
  3. 3.Institute of Environmental Science and Research (ESR) LtdPoriruaNew Zealand

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