Automation of RNA-based biomarker extraction from dried blood spots for the detection of blood doping

Bioanalysis. 2020 Jun;12(11):729-736. doi: 10.4155/bio-2020-0041. Epub 2020 May 15.

Abstract

Aim: Transcriptomic biomarkers originating from reticulocytes measured in dried blood spots (DBSs) may be reliable indicators of blood doping. Methods/results: Here, we examined changes in the expression levels of the erythropoiesis-related ALAS2, CA1 and SLC4A1 genes in DBS samples from elite athletes and volunteers of clinical study with recombinant erythropoietin dose. Conclusion: By comparing the mean intraday coefficients of variation for ALAS2L, ALASLC, CA1 and SLC4A1 between manual and automated RNA extractions, an average improvement was observed, whereas the assessment of interday variability provided comparable results for both manual and automated approaches. Our results confirmed that RNA biomarkers on DBS support are efficient to detect blood doping.

Keywords: RNA extraction; automation; dried blood spot; transcriptomic biomarkers.

MeSH terms

  • 5-Aminolevulinate Synthetase / genetics
  • 5-Aminolevulinate Synthetase / isolation & purification
  • 5-Aminolevulinate Synthetase / metabolism
  • Anion Exchange Protein 1, Erythrocyte / genetics
  • Anion Exchange Protein 1, Erythrocyte / isolation & purification
  • Athletes
  • Automation*
  • Biomarkers / blood
  • Biomarkers / metabolism
  • Carbonic Anhydrase I / genetics
  • Carbonic Anhydrase I / isolation & purification
  • Carbonic Anhydrase I / metabolism
  • Doping in Sports*
  • Dried Blood Spot Testing*
  • Humans
  • RNA / blood*
  • RNA / genetics
  • RNA / isolation & purification
  • Substance Abuse Detection*

Substances

  • Anion Exchange Protein 1, Erythrocyte
  • Biomarkers
  • SLC4A1 protein, human
  • RNA
  • 5-Aminolevulinate Synthetase
  • ALAS2 protein, human
  • Carbonic Anhydrase I