Thrombin generation measurement using the ST Genesia Thrombin Generation System in a cohort of healthy adults: Normal values and variability

Sara Calzavarini, Justine Brodard, Claudia Quarroz, Livia Maire, Raphael Nützi, Jovana Jankovic, Laura C Rotondo, Evelyne Giabbani, Georg Martin Fiedler, Michael Nagler, Anne Angelillo-Scherrer, Sara Calzavarini, Justine Brodard, Claudia Quarroz, Livia Maire, Raphael Nützi, Jovana Jankovic, Laura C Rotondo, Evelyne Giabbani, Georg Martin Fiedler, Michael Nagler, Anne Angelillo-Scherrer

Abstract

Background: Thrombin generation (TG) assays evaluate the balance between pro- and anticoagulant forces, to better assess bleeding and thrombotic risks. Although TG readouts obtained with the calibrated automated TG have been investigated in multiple clinical conditions, TG still needs standardization and clinical validation. The automated TG instrument ST Genesia® (STG, Stago, Asnières-sur-Seine, France) provides a normalization of TG parameters based on a reference plasma aiming to reduce the interlaboratory variability and the variability between different measurement runs.

Objectives: To evaluate STG in a group of healthy adults.

Methods: Reference intervals in healthy adults and variability of the new standardized reagents for bleeding (BleedScreen) and thrombophilic (ThromboScreen) conditions were determined using STG.

Results: TG was measured in platelet-free plasma (PFP) samples of 123 healthy adults. Reference intervals were determined for TG parameters. Intra- and interassay coefficients of variation were calculated on quality controls and PFP samples from healthy adults. Oral contraception (OC) possibly influenced TG parameters, resulting in a higher median and a broader reference interval for peak height and endogenous thrombin potential (ETP) in women aged 20 to 49 years than in all other sex and age categories. Therefore, we propose the following reference interval categories: men, women aged <50 years not using OC, women aged <50 years using OC, and women aged ≥50 years. Normalization was effective to reduce the interassay variability of quality controls for ETP (BleedScreen assay), and peak height and ETP (ThromboScreen assay without thrombomodulin), but had little impact on PFP sample variability.

Conclusion: STG appears suitable for accurate measurement of TG in healthy adults.

Keywords: blood coagulation factors; clinical laboratory tests; quality control; reference ranges; thrombin generation.

© 2019 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals, Inc on behalf of International Society on Thrombosis and Haemostasis.

Figures

Figure 1
Figure 1
ST Genesia BleedScreen parameters in the age categories 20‐49 years and 50‐80 years, respectively. (A) Peak height normalized. (B) Endogenous thrombin potential (ETP) normalized. Data from all men are shown by blue boxes. Data from all women are shown by pink boxes. Data from women aged 20‐49 are divided in 2 groups: women using oral contraception (green boxes) and women not using oral contraception (red boxes). Median and interquartile range are indicated. Groups were compared using the Mann‐Whitney U‐test. The dotted lines show the threshold proposed by Stago. ns, not significant; ****P < 0.0001
Figure 2
Figure 2
ST Genesia ThromboScreen parameters in the age categories 20‐49 and 50‐80, respectively. (A) Peak height normalized. (B) Endogenous thrombin potential (ETP) obtained in absence of thrombomodulin normalized. (C) Endogenous thrombin potential (ETP) obtained in presence of thrombomodulin normalized. Data from all men are shown by blue boxes. Data from all women are shown by pink boxes. Data from women aged 20‐49 are divided in 2 groups: women using oral contraception (green boxes) and women not using oral contraception (red boxes). Medians and interquartile ranges are indicated. Groups were compared using the Mann‐Whitney U‐test. The dotted lines show the threshold proposed by STAGO. TM, thrombomodulin; ns, not significant; **P < 0.01; ***P < 0.001; ****P < 0.0001
Figure 3
Figure 3
Correlations between some hemostasis/thrombophilia parameters and ETP values in the study group. (A) ETP was measured using the BleedScreen assay and the ThromboScreen assay. (B) Endogenous thrombin potential (ETP) inhibition was measured using the ThromboScreen (TS) assay in presence/absence of thrombomodulin. AT, antithrombin; ETP, endogenous thrombin potential; FIX, factor IX; free PS, free protein S; FV, factor V; FVIII, factor VIII; PC:c, protein C coagulant; r, Pearson coefficient; STG‐BLS, BleedScreen; STG‐TS, ThromboScreen

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