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mRNA Biomarkers in Dried Blood Spots May Improve Detection of Autologous Blood Micro-Transfusions Using an
Andreas Breenfeldt Andersen1, Jessica Almeida Oliveira2, Francesco Loria2
1Department of Public Health, Research Unit for Exercise Biology, Aarhus University, Aarhus, Denmark.
Drug Testing and Analysis
|August 9, 2025
Summary
Detecting micro-doses of autologous blood transfusions (ABMTs) is challenging. Measuring ALAS2 mRNA from dried blood spots shows promise as a sensitive biomarker for identifying ABMTs, enhancing anti-doping efforts.
Area of Science:
- Sports Science
- Biochemistry
- Anti-Doping Research
Background:
- Autologous blood transfusions (ABTs) are prohibited by the World Anti-Doping Agency (WADA).
- Detecting smaller volume autologous blood micro-transfusions (ABMTs) is difficult due to attenuated biomarker changes.
- Current Athlete Biological Passport (ABP) methods lack sensitivity for detecting ABMTs.
Purpose of the Study:
- To evaluate mRNA expression of 5-aminolevulinic acid synthase (ALAS2) and carbonic anhydrase 1 (CA1) from dried blood spots (DBS) as potential biomarkers for ABMT detection.
- To assess the sensitivity and specificity of these mRNA biomarkers in trained individuals undergoing a controlled ABMT protocol.
Main Methods:
- A randomized, placebo-controlled study involving 47 trained individuals.
- ABMT group received a 130 mL packed red blood cell reinfusion 4 weeks after blood donation.
- Analysis of ALAS2 and CA1 mRNA expression from DBS, and reticulocyte percentage (RET%) from venous blood, before and after donation/reinfusion.
Main Results:
- Significant increases in ALAS2, CA1 mRNA, and RET% were observed after blood donation.
- No consistent group-level changes were detected immediately after ABMT.
- Individualized analysis of ALAS2 mRNA expression demonstrated >95% sensitivity and specificity for ABMT detection.
Conclusions:
- ALAS2 mRNA expression measured from DBS is a sensitive and specific biomarker for detecting ABMT.
- This minimally invasive approach using DBS sampling can potentially improve anti-doping strategies against small-volume autologous transfusions.
- ALAS2 mRNA analysis offers a promising tool to enhance the detection capabilities beyond traditional ABP markers.
