We use cookies to improve your experience. By continuing to browse this site, you accept our cookie policy.×
Skip main navigation
Open Drawer MenuClose Drawer Menu
Bioanalysis
BioTechniques
Future Drug Discovery
Future Medicinal Chemistry
Future Science OA
International Journal of Pharmacokinetics
Pharmaceutical Patent Analyst
Therapeutic Delivery
Preliminary Communication

Monitoring of hemoglobin and erythropoiesis-related mRNA with dried blood spots in athletes and patients

    Francesco Loria

    Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne & Geneva, Lausanne University Hospital & University of Lausanne, Switzerland

    Search for more papers by this author

    ,
    Andreas P Stutz

    Service of Nephrology, Lausanne University Hospital & University of Lausanne, Lausanne, Switzerland

    Search for more papers by this author

    ,
    Angela Rocca

    Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne & Geneva, Lausanne University Hospital & University of Lausanne, Switzerland

    Search for more papers by this author

    ,
    Silke Grabherr

    University Center of Legal Medicine, Lausanne & Geneva, Lausanne University Hospital & University of Lausanne, Switzerland

    Search for more papers by this author

    ,
    Tiia Kuuranne

    Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne & Geneva, Lausanne University Hospital & University of Lausanne, Switzerland

    Search for more papers by this author

    ,
    Menno Pruijm

    Service of Nephrology, Lausanne University Hospital & University of Lausanne, Lausanne, Switzerland

    Search for more papers by this author

    &
    Nicolas Leuenberger

    *Author for correspondence: Tel.: +41 21 31470 95;

    E-mail Address: Nicolas.leuenberger@chuv.ch

    Swiss Laboratory for Doping Analyses, University Center of Legal Medicine, Lausanne & Geneva, Lausanne University Hospital & University of Lausanne, Switzerland

    Search for more papers by this author

    Published Online:17 Feb 2022https://doi.org/10.4155/bio-2021-0252

    Abstract

    Aim: We assessed the feasibility of using hematological parameters (such as hemoglobin and reticulocyte mRNA) in dried blood spot (DBS) samples to test athletes for doping and to improve patient care. Methods: Hemoglobin and erythropoiesis-related mRNAs were measured in venous blood and DBSs from both healthy athletes and hemodialysis patients. Results: We accurately measured hemoglobin changes over time in both venous blood and DBS samples. Combining hemoglobin and mRNA analyses, we detected erythropoietin injection in DBSs more sensitively and with higher efficiency by using the DBS OFF-score than by using the athlete biological passport OFF-score. Conclusion: DBS-based measurements are practical for calculating hemoglobin levels and athlete biological passport OFF-scores. This approach may help detect blood doping and help predict patient response to EPO.

    Papers of special note have been highlighted as: • of interest; •• of considerable interest

    References

    • 1. World Anti-Doping Agency. WADA technical document: analytical requirements for the hematological module of the athlete biological passport (TD2021Bar) (2021). www.wada-ama.org/en/resources/lab-documents/td2021bar
      Google Scholar
    • 2. Tuck MK, Chan DW, Chia D et al. Standard operating procedures for serum and plasma collection: Early Detection Research Network Consensus Statement Standard Operating Procedure Integration Working Group. J. Proteome Res. 8(1), 113–117 (2009).
      Medline CASGoogle Scholar
    • 3. Sharma A, Jaiswal S, Shukla M, Lal J. Dried blood spots: concepts, present status, and future perspectives in bioanalysis: an overview on dried blood spots. Drug Test. Anal. 6(5), 399–414 (2014). • Describes the concept and the benefits of dried blood spots.
      Medline CASGoogle Scholar
    • 4. Reverter-Branchat G, Ventura R, Ezzel Din M, Mateus J, Pedro C, Segura J. Detection of erythropoiesis-stimulating agents in a single dried blood spot. Drug Test. Anal. 10(10), 1496–1507 (2018).
      Medline CASGoogle Scholar
    • 5. Rocca A, Martin L, Kuuranne T, Ericsson M, Marchand A, Leuenberger N. A fast screening method for the detection of CERA in dried blood spots. Drug Test. Anal. doi: 10.1002/dta.3142 (2021) (Epub ahead of print). • First paper demonstrating the possibility to detect CERA in dried blood spots.
      Google Scholar
    • 6. Cox HD, Miller GD, Lai A, Cushman D, Eichner D. Detection of autologous blood transfusions using a novel dried blood spot method. Drug Test. Anal. 9(11–12), 1713–1720 (2017).
      Medline CASGoogle Scholar
    • 7. Salamin O, Gottardo E, Schobinger C et al. Detection of stimulated erythropoiesis by the RNA-based 5′-aminolevulinate synthase 2 biomarker in dried blood spot samples. Clin. Chem. 65(12), 1563–1571 (2019). •• First paper demonstrating longitudinal follow-up of RNA-based biomarkers to detect blood doping on dried blood spots.
      Medline CASGoogle Scholar
    • 8. Loria F, Manfredi M, Reverter-Branchat G, Segura J, Kuuranne T, Leuenberger N. Automation of RNA-based biomarker extraction from dried blood spots for the detection of blood doping. Bioanalysis 12(11), 729–736 (2020). •• First paper demonstrating automated extraction of mRNA from dried blood spots.
      CASGoogle Scholar
    • 9. Loria F, Cox HD, Voss SC et al. The use of RNA-based 5′-aminolevulinate synthase 2 biomarkers in dried blood spots to detect recombinant human erythropoietin microdoses. Drug Test. Anal. doi: 10.1002/dta.3123 (2021) (Epub ahead of print). •• Showed the effect of hypoxia and micro-doses of recombinant human erythropoietin injection on reticulocyte-related mRNA.
      MedlineGoogle Scholar
    • 10. Richardson G, Marshall D, Keevil B. Prediction of haematocrit in dried blood spots from the measurement of haemoglobin using commercially available sodium lauryl sulphate. Ann. Clin. Biochem. 55(3), 363–367 (2018).
      Medline CASGoogle Scholar
    • 11. Oshiro I, Takenaka T, Maeda J. New method for hemoglobin determination by using sodium lauryl sulfate (SLS). Clin. Biochem. 15(2), 83–88 (1982).
      Medline CASGoogle Scholar
    • 12. Thevis M. Anti-doping analysis. Anal. Bioanal. Chem. 401(2), 387–388 (2011).
      Medline CASGoogle Scholar
    • 13. Babitt JL, Lin HY. Mechanisms of anemia in CKD. JASN 23(10), 1631–1634 (2012).
      Medline CASGoogle Scholar
    • 14. Collister D, Rigatto C, Tangri N. Anemia management in chronic kidney disease and dialysis: a narrative review. Curr. Opin. Nephrol. Hypertens. 26(3), 214–218 (2017).
      MedlineGoogle Scholar
    • 15. Sharpe K, Ashenden MJ, Schumacher YO. A third generation approach to detect erythropoietin abuse in athletes. Haematologica 91(3), 356–363 (2006).
      Medline CASGoogle Scholar
    • 16. World Anti-Doping Agency. WADA technical document to harmonize the detection and reporting of erythropoietin (EPO) and other EPO-receptor agonists (ERAs) by laboratories when analyzed using polyacrylamide gel-electrophoretic (PAGE) analytical methods (TD2021EPO) (2022). www.wada-ama.org/en/resources/lab-documents/td2022epo
      Google Scholar
    • 17. Capiau S, Stove VV, Lambert WE, Stove CP. Prediction of the hematocrit of dried blood spots via potassium measurement on a routine clinical chemistry analyzer. Anal. Chem. 85(1), 404–410 (2013).
      Medline CASGoogle Scholar
    • 18. Capiau S, Wilk LS, Aalders MCG, Stove CP. A novel, nondestructive, dried blood spot-based hematocrit prediction method using noncontact diffuse reflectance spectroscopy. Anal. Chem. 88(12), 6538–6546 (2016).
      Medline CASGoogle Scholar
    • 19. Salamin O, Mignot J, Kuuranne T, Saugy M, Leuenberger N. Transcriptomic biomarkers of altered erythropoiesis to detect autologous blood transfusion. Drug Test. Anal. 10(3), 604–608 (2018).
      Medline CASGoogle Scholar