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Impact of Centrifugation on Thrombin Generation Analysis Evaluated in Healthy Subjects and Patients Treated with Direct Oral Anticoagulants

1Université Catholique de Louvain, CHU UCL Namur, Hematology Laboratory, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), Yvoir, Belgium, 2CHU UCL Namur, Department of Anaesthesiology, Namur Thrombosis and Haemostasis Centre (NTHC), Namur Research Institute for Life Sciences (NARILIS), Université catholique de Louvain, Yvoir, Belgium, 3Ospedale di Cremona, Hemostasis and Thrombosis Center, Cremona, Italy, 4University of Namur, Department of Pharmacy, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), Namur, Belgium, 5Qualiblood s.a., Namur, Belgium, 6Diagnostica Stago, Asnières-sur-Seine, France

Abstract Number: PB0634

Meeting: ISTH 2020 Congress

Theme: Diagnostics and OMICs » Laboratory Diagnostics

Background: Double centrifugation before freezing is recommended for thrombin generation (TG) analysis. However, this preanalytical procedure is often time consuming and not well managed in clinical practice.

Aims: The aim of this study was to evaluate the impact of single and double centrifugation on TG results measured on frozen samples of healthy volunteers (HV) and patients treated with direct oral anticoagulants (DOAC).

Methods: Forty HV and 57 patients receiving a DOAC (apixaban, dabigatran, edoxaban or rivaroxaban) were included in this prospective double-center observational study. Blood was collected in 109mM citrated tubes and frozen at minimum -70°C before TG measurement using ST-Genesia® with STG-DrugScreen® reagent. Four pre-analytical conditions were studied: (A) single centrifugation (2000g, 15min) before freezing; (B) one centrifugation before freezing and another after thawing (2000g, 15min for both); (C) one centrifugation before freezing and another after thawing (2000g, 15 min then 2000g, 10 min); (D) double centrifugation (2000g, 15min) before freezing (reference).
Normalized lag time (LTn), endogenous thrombin potential (ETPn) and peak height (PHn) were compared using the reference condition. Acceptable relative differences were defined at 6%, 8% and 10% for LTn, ETPn and PHn, respectively.

Results: TG parameters were significantly different between the two sites for dabigatran patients and for HV. Centrifugation conditions had no clinically significant impact on HV but single centrifugation always resulted in clinically significant reductions in LTn for DOAC samples (Figure 1). A second centrifugation after thawing permits the recovery of clinically acceptable differences for the three TG parameters for edoxaban and dabigatran (site 1) samples, but not for apixaban, rivaroxaban nor dabigatran (site 2) samples (Table 1).

Conclusions: Overall, single centrifugation before freezing is acceptable only for edoxaban samples and only if a second centrifugation is performed after thawing. Double centrifugation before freezing should remain the recommended pre-analytical condition for TG measurement.

    Apixaban Edoxaban Rivaroxaban Dabigatran Site 1 Dabigatran Site 2 Healthy volunteers Site 1 Healthy volunteers Site 2
  A vs. D -6.5% ** -11.1% ** -11.2% *** -22.5% * -20.9% ** -3.7% ** -5.9% ***
LTn B vs. D -7.9% ** -5.8% ** -7.9% *** -12.5% -10.1% ** -3.5% ** -4.1% ***
  C vs. D -6.6% * -9.2% -6.8% ** -25.6% * -12.3% ** -4.0% ** -5.7% ***
  A vs. D +14.0% *** +9.1% ** +13.0% *** +3.9% +1.6% +2.7% * +1.2% *
PHn B vs. D +14.2% ** +2.2% +3.0% +4.8% +2.3% * +3.9% ** +2.8% ***
  C vs. D +14.2% ** +9.6% +7.3% ** +7.8% *** +2.3% * +2.8% * +2.3% ***
  A vs. D +8.4% * +6.1% * +15.1% *** +3.3% -1.7% +1.7% +1.5% *
ETPn B vs. D +4.2% * +1.2% +2.2% -3.7% +0.3% +1.6% +1.5% *
  C vs. D +5.4% * +2.6% +7.5% * +1.4% +0.8% +1.5% +1.1%

[Differences of normalized thrombin generation parameters between pre-analytical conditions A, B, C and D (see text for description).]


[Normalized lag time (LTn) in the different centrifugation conditions (A, B, C and D; see text) for each DOAC.]

To cite this abstract in AMA style:

Hardy M, Dellanoce C, Douxfils J, Sarah L, Carlo A, Testa S, Mullier F. Impact of Centrifugation on Thrombin Generation Analysis Evaluated in Healthy Subjects and Patients Treated with Direct Oral Anticoagulants [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/impact-of-centrifugation-on-thrombin-generation-analysis-evaluated-in-healthy-subjects-and-patients-treated-with-direct-oral-anticoagulants/. Accessed September 21, 2023.

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