Dual-color bioluminescent bioreporter for forensic analysis: evidence of androgenic and anti-androgenic activity of illicit drugs

Anal Bioanal Chem. 2013 Jan;405(2-3):1035-45. doi: 10.1007/s00216-012-6416-6. Epub 2012 Sep 26.


Bioassays represent promising complementary techniques to conventional analytical approaches used in doping analysis to detect illicit drugs like anabolic-androgenic steroids (AAS). The fact that all AAS share a common mechanism of action via the human androgen receptor (hAR) enables the use of bioassays, relying on the activation of hAR as antidoping screening tools. Previously, we developed a dual-color bioreporter based on yeast cells engineered to express hAR and androgen response elements driving the expression of the bioluminescent (BL) reporter protein Photinus pyralis luciferase. A second reporter protein, the red-emitting luciferase PpyRE8, was introduced in the bioreporter as internal viability control. Here, we report the first forensic application of a straightforward, accurate, and cost-effective bioassay, relying on spectral resolution of the two BL signals, in 96-microwell format. The bioreporter responds to dihydrotestosterone as reference androgen in a concentration-dependent manner from 0.08 to 1,000 nM with intra- and inter-assay variation coefficients of 11.4 % and 13.1 %, respectively. We also demonstrated the suitability of this dual-color bioreporter to assess (anti)-androgenic activity of pure AAS, mixtures of AAS, and other illicit drugs provided by the Scientific Police. Significant anti-androgenic activity was observed in samples labeled as marijuana and hashish, containing Δ(9)-tetrahydrocannabinol as major constituent.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androgen Antagonists / analysis*
  • Androgen Antagonists / urine
  • Androgens / analysis*
  • Androgens / urine
  • Biosensing Techniques / methods*
  • Forensic Medicine / methods*
  • Genes, Reporter
  • Humans
  • Illicit Drugs / analysis*
  • Illicit Drugs / urine
  • Luciferases / analysis
  • Luciferases / urine
  • Luminescent Measurements / methods*
  • Male
  • Receptors, Androgen / genetics
  • Receptors, Androgen / metabolism
  • Steroids / analysis
  • Steroids / urine


  • Androgen Antagonists
  • Androgens
  • Illicit Drugs
  • Receptors, Androgen
  • Steroids
  • Luciferases