Detection of autologous blood transfusions using a novel dried blood spot method

Abstract

In doping control laboratories, autologous blood transfusions are currently detected using an indirect method that monitors changes in an athlete's hemoglobin concentration [Hb] and reticulocyte percent (Ret%) over time. The method is limited by the need for a phlebotomist to collect venous blood and the limited blood stability which requires temperature-controlled shipment and analysis within 72 hours. These limitations significantly reduce the number of samples collected from each athlete and thus the utility of the method. We have recently developed a method to measure immature reticulocytes (IRC) and red blood cells (RBC) in dried blood spots, which could replace the current venous blood method. In the DBS method, cell-specific proteins are digested with trypsin and measured by mass spectrometry. Two proteins, CD71 and Band3, are measured to count IRC and RBC, respectively. The method was tested in an autologous transfusion study consisting of 15 subjects who received blood and 11 subjects who received saline. After transfusion, the average CD71/Band3 ratio in the blood group was statistically different from the saline group at days 5, 6, 13, and 20. The average CD71/Band3 ratio decreased to a minimum of 61 ± 8% of baseline, while Ret% decreased to 75 ± 5% of baseline. Based on experimentally defined criteria, the CD71/Band3 ratio could detect 7 out of 10 blood transfusion subjects, while Ret% could detect 3 out of 10. Thus, the DBS method could improve detection of autologous transfusion and allow increased sample collection.

Can dried blood spots (DBS) contribute to conducting comprehensive SARS‐CoV‐2 antibody tests?

“…A proof-of-principle pilot study was conducted to probe for options as to how routine sports drug testing programs can contribute to generating information on the prevalence of SARS-CoV-2 antibodies in the sub-population of elite athletes. As DBS tests have been studied in the context of general doping controls extensively over the past decade, [16][17][18][19][20][21][22][23][24][25] the applicability of two commercially available IgG and IgM tests, one well-plate ELISA assay and one lateral flow-based rapid test, to the analysis of paired DBS and blood plasma samples obtained from 26 individuals was assessed.…”

Analysis of insulin and insulin analogs from dried blood spots by means of liquid chromatography–high resolution mass spectrometry

“…Most of the existing methods for lower and intermediate molecular mass peptide hormones are based on urine or blood analysis accordingly. But also methods for DBS analysis of small molecules as well as selected peptide hormones in doping controls were published and the implementation of this alternative sample collection is expected in the near future . Thus, analytical methods to cover the entire portfolio of prohibited substances is required for DBS analysis.…”

Annual banned‐substance review: Analytical approaches in human sports drug testing

“…Another alternative test method relying on DBS rather than whole blood was presented by Cox et al, who determined the ratio of immature reticulocytes and red blood cells via their cell‐specific proteins CD71 and Band3, respectively . In a pilot study, 15 participants underwent autologous blood transfusion, and CD71/Band3 ratios were compared to a saline transfused control group ( n = 11).…”