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Comparison of Dried Blood Spot Sampling Methods for RNA-Based Biomarker Measurement in Anti-Doping.

Jacob Bejder1, Corentin Schlechten2, Claudia Mumenthaler3

  • 1Department of Nutrition, Exercise and Sports (NEXS), University of Copenhagen, Copenhagen, Denmark.

Drug Testing and Analysis
|May 20, 2026
PubMed
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Dried blood spot (DBS) sampling methods provide comparable results for anti-doping RNA biomarkers. This allows for flexible, less invasive monitoring in sports doping control and research.

Area of Science:

  • Biomarker analysis
  • Doping control
  • Molecular diagnostics

Background:

  • Dried blood spot (DBS) sampling offers logistical benefits over traditional blood collection.
  • The comparability of analytical results across different DBS collection methods remains underexplored.
  • Investigating DBS method agreement is crucial for anti-doping and longitudinal biomarker monitoring.

Purpose of the Study:

  • To assess the comparability of RNA-based erythropoiesis biomarkers (ALAS2, CA1) across different DBS sampling methods.
  • To evaluate venous vs. capillary blood collection on cellulose cards and the Tasso-M20 device.
  • To determine suitability for anti-doping and research applications requiring decentralized sampling.

Main Methods:

  • Collected DBS samples from athletes and healthy volunteers using venous-cellulose, capillary-cellulose, and Tasso-M20 methods.
Keywords:
ALAS2CA1RNA biomarkersTasso‐M20anti‐dopingdried blood samplingerythropoietin

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  • Quantified ALAS2 and CA1 mRNA expression using quantitative reverse transcription PCR (RT-qPCR).
  • Assessed agreement between matrices using Passing-Bablok regression analysis.

    • Main Results:

    • Strong linear agreement (r ≥ 0.96) observed for ALAS2 and CA1 expression across all tested DBS matrices.
    • ALAS2 showed no constant or proportional bias across comparisons; CA1 exhibited a minor proportional bias (~9-22%) in one comparison.
    • No significant differences in relative RNA expression were found across matrices during a recombinant human erythropoietin (rEPO) administration study.

    Conclusions:

    • ALAS2 and CA1 mRNA can be reliably quantified across various DBS sampling methods (cellulose and Tasso-M20).
    • The observed minimal bias for CA1 is acceptable relative to biological variability, supporting longitudinal monitoring.
    • These findings validate DBS for less invasive, flexible, and decentralized anti-doping and research sampling strategies.