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Sensitivity and specificity of presumptive tests for blood, saliva and semen



Despite their wide use, the limits of presumptive tests can be poorly understood. The aim of this study was to investigate the specificity and sensitivity of conventional, as well as innovative, presumptive tests for blood, semen and saliva.


We investigated Kastle–Meyer (KM) and leucomalachite green (LMG) tests for blood with regard to their sensitivity and specificity in the presence of oxidizing (hypochlorite) and anti-oxidizing (ascorbic acid) agents. The suitability and specificity of the red starch paper (RSP) test for saliva was assessed. Finally, the inhibitory effect of detergent on the acid phosphatase (AP) test for semen was investigated along with possible cross reactions to tea stains.


Our results confirm previous findings of higher sensitivity and specificity of the KM test compared to LMG test for blood. Contrary to previous studies, no statistically significant difference was observed in the sensitivity of the tests between dry and wet stains. The novel RSP test was found to successfully detect saliva. We demonstrated that acid phosphatase (AP) testing for semen is possible on used RSP. A common multipurpose detergent had an inhibitory effect on AP tests. False positive results were obtained from tea stains. Testing different sorts of tea (black, green and herbal teas) revealed that only Camellia varieties produce positive result with the AP test, due to AP being present in the plants.


From our results we conclude that specific knowledge of each test, including substances that may affect the test outcome, is imperative to ensure correct interpretation of presumptive test results.

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  1. Kastle JH, Shedd OM. Phenolphthalin as a reagent for the oxidizing ferments. Am Chem J. 1901;26:526.

    CAS  Google Scholar 

  2. Meyer E. Beiträge zur Leukocytenfrage. Münchner Medizinische Wochenschrift. 1903;50:1489–93.

    Google Scholar 

  3. Cox M. A study of the sensitivity and specificity of four presumptive tests for blood. J Forensic Sci. 1991;36:1503–11.

    CAS  PubMed  Google Scholar 

  4. Kind SS. The use of the acid phosphatase test in searching for seminal stains. J Crim Law Criminol Police Sci. 1957;47:597–600.

    CAS  Article  Google Scholar 

  5. Tobe SS, Watson N, Daéid NN. Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA. J Forensic Sci. 2007;52:102–9.

    CAS  PubMed  Article  Google Scholar 

  6. Grodsky M, Wright K, Kirk PL. Simplified preliminary blood testing-an improved technique and a comparative study of methods. J Crim Law Criminol Police Sci. 1951;42:95–104.

    CAS  Article  Google Scholar 

  7. Hooft PJ, van de Voorde HP. Interference of body products, food and products from daily life with the modified zinc test and the acid phosphatase test. Forensic Sci Int. 1994;66:187–96.

    CAS  PubMed  Article  Google Scholar 

  8. Tirimanna AS. Acid phosphatases in the tea leaf. Experientia. 1972;28:633.

    CAS  PubMed  Article  Google Scholar 

  9. Martin NC, Clayson NJ, Scrimger DG. The sensitivity and specificity of red-starch paper for the detection of saliva. Sci Justice. 2006;46:97–105.

    CAS  PubMed  Article  Google Scholar 

  10. Hedman J, Dalin E, Rasmusson B, Ansell R. Evaluation of amylase testing as a tool for saliva screening of crime scene trace swabs. Forensic Sci Int Genet. 2011;5:194–8.

    CAS  PubMed  Article  Google Scholar 

  11. Bright JA, Cockerton S, Harbison S, Russell A, Samson O, Stevenson K. The effect of cleaning agents on the ability to obtain DNA profiles using the Identifiler™ and PowerPlex® Y multiplex kits. J Forensic Sci. 2011;56:181–5.

    PubMed  Article  Google Scholar 

  12. Sackiewicz A, Niemcunowicz-Janica A, Pepiński W, Skawrońska M, Szeremeta M, Ptaszyńska-Sarosiek I, Okłota M. Evaluation of visualization of biological stains with the use of alternative light source (ALS) for the purpose of genetic identification. Part II. Semen samples analysis. Arch Med Sadowej Kryminol. 2010;60:258–62.

    PubMed  Google Scholar 

  13. Haas C, Hanson E, Ballantyne J. Capillary electrophoresis of a multiplex reverse transcription-polymerase chain reaction to target messenger RNA markers for body fluid identification. Methods Mol Biol. 2012;830:169–83.

    CAS  PubMed  Article  Google Scholar 

  14. Gršković B, Zrnec D, Vicković S, Popović M, Mršić G. DNA methylation: the future of crime scene investigation? Mol Biol Rep. 2013;40:4349–460.

    PubMed  Article  Google Scholar 

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The authors thank all students of the Forensic Science postgraduate classes of 2010/2011 and 2011/2012 for their outstanding laboratory work. We also thank Nicola McCallum for excellent technical assistance.

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Correspondence to Marielle Vennemann.

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Vennemann, M., Scott, G., Curran, L. et al. Sensitivity and specificity of presumptive tests for blood, saliva and semen. Forensic Sci Med Pathol 10, 69–75 (2014).

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  • Biological presumptive test
  • Sensitivity
  • Specificity
  • Acid phosphatase
  • Tea leaves