Validation of a Blood Stability Score as an easy‐to‐use blood sample quality index

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.

Section: Introductionmentioning

confidence: 99%

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

Cox¹,

Miller

Lai³

et al. 2017

“…Ten total baseline samples from 10 subjects, collected 2 days before the event, were excluded from the study. According to the Guidelines provided by WADA, the ABP blood sample validity is dependent on a calculated blood stability score, which factors elapsed time from collection to analysis as well as the mean shipping temperature . As this particular sample shipment was delayed, the collection‐to‐analysis time surpassed the allowable limit and thus the samples were invalidated and not included in the data analysis for this study.…”

Section: Resultsmentioning

confidence: 99%

“…Significance was determined using a one-way ANOVA with Dunnett's multiple comparisons test, comparing each post-race day against the baseline calculated blood stability score, which factors elapsed time from collection to analysis as well as the mean shipping temperature. 15 As this particular sample shipment was delayed, the collection-to-analysis time surpassed the allowable limit and thus the samples were invalidated and not included in the data analysis for this study. As a result, some of the subjects in this study did not present with three total baseline samples.…”

Section: Exclusion Of Samplesmentioning

confidence: 99%

Hematological changes following an Ironman triathlon: An antidoping perspective

Miller¹,

Beharry

Teramoto

et al. 2019

Understanding and characterizing confounding factors to the Athlete Biological Passport (ABP) is crucial for the reliable interpretation of biological profiles in the antidoping field. The physiological effects on hematological parameters and plasma volume (PV) following competition in a long‐distance triathlon, as seen in the Ironman discipline, have yet to be fully described and are the focus of this study. Complete blood count blood tests were conducted on 19 Ironman triathletes before and after an Ironman triathlon to characterize changes in hematological parameters and the effect on ABP interpretation, as it was hypothesized that changes in the plasma volume may result in the presentation of atypical ABP profiles. Baseline blood samples were collected from the athletes prior to the event, and one sample was collected per day for up to 1 week following the race. Differences were observed between the male and female athletes across multiple parameters. Most importantly to the ABP, decreases in hemoglobin concentration (HGB) and hematocrit (HCT) were identified post‐race, with the largest decreases identified on day +2. The average HGB returned to pre‐race baseline levels on day +5. Beginning 5–6 days after the race, increases in the reticulocyte percentage (Ret%) were identified. Atypical Passport Findings were identified in 32% (6/19) of the ABPs, flagged mainly due to atypical hemoglobin concentration and one instance in which the OFF‐score exceeded the adaptive model limits. These results offer a characterized timeline of hematological changes and expected shifting of plasma volume following an Ironman triathlon providing important data for the reliable interpretation of ABP profiles in this field.

“…Producing ABP data in routine doping controls necessitates frequent blood sampling, which is associated with significant costs arising from sampling and transport requirements. The blood sample quality must not be compromised, and a blood stability score (BSS) was suggested by Robinson et al that should determine the validity of a blood specimen upon arrival in the doping control laboratory . Taking into account the time period between sample collection and analysis (CAT) and the storage temperature (T) of the sample, the BSS was defined by BSS = CAT + 3xT, and values below 85 were found to guarantee the stability of parameters relevant for ABP analyses.…”

Section: Manipulation Of Blood and Blood Componentsmentioning

confidence: 99%

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

Thevis

Kuuranne

Geyer

2017

Several high-profile revelations concerning anti-doping rule violations over the past 12 months have outlined the importance of tackling prevailing challenges and reducing the limitations of the current anti-doping system. At this time, the necessity to enhance, expand, and improve analytical test methods in response to the substances outlined in the World Anti-Doping Agency's (WADA) Prohibited List represents an increasingly crucial task for modern sports drug-testing programs. The ability to improve analytical testing methods often relies on the expedient application of novel information regarding superior target analytes for sports drug-testing assays, drug elimination profiles, alternative test matrices, together with recent advances in instrumental developments. This annual banned-substance review evaluates literature published between October 2016 and September 2017 offering an in-depth evaluation of developments in these arenas and their potential application to substances reported in WADA's 2017 Prohibited List.