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mRNA Biomarkers in Dried Blood Spots May Improve Detection of Autologous Blood Micro-Transfusions Using an Individualized Approach

Title: mRNA Biomarkers in Dried Blood Spots May Improve Detection of Autologous Blood Micro-Transfusions Using an Individualized Approach
Authors: Andersen, Andreas Breenfeldt; Oliveira, Jessica Almeida; Loria, Francesco; Bejder, Jacob; Salamin, Olivier; Kuuranne, Tiia; Nordsborg, Nikolai B; Leuenberger, Nicolas
Source: Andersen, AB, Oliveira, JA, Loria, F, Bejder, J, Salamin, O, Kuuranne, T, Nordsborg, NB & Leuenberger, N 2025, 'mRNA Biomarkers in Dried Blood Spots May Improve Detection of Autologous Blood Micro-Transfusions Using an Individualized Approach', Drug Testing and Analysis, vol. 17, no. 11, pp. 2291-2300. https://doi.org/10.1002/dta.3939
Publication Year: 2025
Collection: Aarhus University: Research
Description: Autologous blood transfusions (ABTs) are prohibited by the World Anti-Doping Agency (WADA), yet detecting autologous blood micro-transfusions (ABMTs) remains a challenge. Due to smaller transfused volumes, ABMTs cause attenuated biomarker changes, limiting detection sensitivity within the Athlete Biological Passport (ABP). This study assessed whether mRNA expression of 5-aminolevulinic acid synthase (ALAS2) and carbonic anhydrase 1 (CA1), measured from dried blood spots (DBS), could serve as sensitive biomarkers of ABMT. In a randomized, placebo-controlled design, 47 trained individuals (24 ♀; mean VO 2 peak 56 ± 7 mL·min −1 ·kg −1 ) were allocated to an ABMT group (n = 23; ♀ = 12) or placebo group (n = 24; ♀ = 12). The ABMT group donated 450 mL of blood and received a 130 mL packed red blood cell reinfusion 4 weeks later. Blood sampling occurred regularly before and after both donation and reinfusion. ALAS2 and CA1 mRNA expression from DBS, and reticulocyte percentage (RET%) from venous blood, were analyzed. Following blood donation, ALAS2, CA1, and RET% increased by 270%, 200%, and 150%, respectively. However, no consistent group-level changes were observed after ABMT. Individualized analysis identified more outliers for ALAS2 than for CA1, and blinded interpretation of individual mRNA profiles achieved > 95% sensitivity and specificity for detecting ABMT. These findings suggest that ALAS2 mRNA expression, assessed via minimally invasive DBS sampling, is a promising biomarker for identifying ABMT. This approach may enhance current anti-doping strategies by improving sensitivity to small-volume autologous transfusions that evade detection through traditional ABP biomarkers.
Document Type: article in journal/newspaper
Language: English
ISSN: 1942-7603; 1942-7611
Relation: info:eu-repo/semantics/altIdentifier/pmid/40781904; info:eu-repo/semantics/altIdentifier/pissn/1942-7603; info:eu-repo/semantics/altIdentifier/eissn/1942-7611
DOI: 10.1002/dta.3939
Availability: https://pure.au.dk/portal/en/publications/20cb3c09-62ff-4eaf-82ca-fc65989b34ff; https://doi.org/10.1002/dta.3939; https://www.scopus.com/pages/publications/105012830952
Rights: info:eu-repo/semantics/openAccess
Accession Number: edsbas.14521F12
Database: BASE