Assessment of the analytical performances and sample stability on ST Genesia system using the STG‐DrugScreen application

Douxfils, Jonathan; Morimont, Laure; Bouvy, Céline; Saint-Hubert, Marie de; Devalet, Bérangère; Devroye, Célia; Dincq, Anne-Sophie; Dogné, Jean‐Michel; Guldenpfennig, Maïté; Baudar, Justine; Larock, Anne-Sophie; Lessire, Sarah; Mullier, François

doi:10.1111/jth.14470

Cited by 44 publications

(88 citation statements)

References 36 publications

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“…The choice of using a commercial reference plasma is in line with the literature since it has been reported that the use of local plasma (i.e., a reference plasma performed at the laboratory facilities) was generally unable to improve the inter‐laboratory variability and the CV is even worsened after normalization . Thus, as mentioned in a previous work and confirmed in this larger series of experiments, the use of a commercial reference plasma is probably useful for the normalization of the TGT parameters such as ETP, peak, lag time, or time‐to‐peak, but not for the calculation of the nAPCsr …”

Section: Discussionsupporting

confidence: 57%

“…27,[43][44][45] The choice of using a commercial reference plasma is in line with the literature since it has been reported that the use of local plasma (i.e., a reference plasma performed at the laboratory facilities) was generally unable to improve the inter-laboratory variability and the CV is even worsened after normalization. 28,43,45 Thus, as mentioned in a previous work 46 and confirmed in this larger series of experiments, the use of a commercial reference plasma is probably useful for the normalization of the TGT parameters such as ETP, peak, lag time, or time-to-peak, but not for the calculation of the nAPCsr. 46,47 Thus, even if it is out of the scope of this study, the reasons of using a reference plasma relies on the fact that the TGT data generated in a single run (i.e., the analysis of the results in absence of added APC) could latter serve in algorithm to assess the prothrombotic tendency of a plasma sample and thus, the between-run normalization of the TGT parameters is of interest in this aspect.…”

Section: Reference Plasma: Which One To Use and To Normalize What?supporting

confidence: 51%

“…28,43,45 Thus, as mentioned in a previous work 46 and confirmed in this larger series of experiments, the use of a commercial reference plasma is probably useful for the normalization of the TGT parameters such as ETP, peak, lag time, or time-to-peak, but not for the calculation of the nAPCsr. 46,47 Thus, even if it is out of the scope of this study, the reasons of using a reference plasma relies on the fact that the TGT data generated in a single run (i.e., the analysis of the results in absence of added APC) could latter serve in algorithm to assess the prothrombotic tendency of a plasma sample and thus, the between-run normalization of the TGT parameters is of interest in this aspect. 48,49 Such normalization has been reported to be better obtained using commercial reference plasma instead of in-house reference plasma.…”

Section: Reference Plasma: Which One To Use and To Normalize What?supporting

confidence: 51%

“…Thus, the solution is to compare the response of the commercial reference plasma with a smaller pool of plasma from healthy donors collected in ideal conditions (i.e., our in‐house reference plasma) and then to apply a correction factor to the commercial plasma to compensate for these differences. This approach has already been proven to be efficient in this and other setting …”

Section: Discussionmentioning

confidence: 99%

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Proof of concept of a new scale for the harmonization and the standardization of the ETP‐based APC resistance

Morimont

Bouvy²,

Delvigne³

et al. 2020

Journal of Thrombosis and Haemostasis

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Background The evaluation of the activated protein C resistance (APCr) based on the endogenous thrombin potential (ETP) is recommended during the development of steroid contraceptives. Results are usually expressed as “normalized APC sensitivity ratio” (nAPCsr) using a reference plasma that should achieve an ETP ratio of 0.1 in presence of exogenous APC. Because of the interassay variability, achieving exactly an ETP ratio of 0.1 in each run is almost impossible, which significantly affects the theoretical 0‐10 scale of nAPCsr. Objectives To compare the nAPCsr to the nAPCsr10, a newly proposed method to express the degree of APC resistance. Methods Individual plasma samples (n = 854) were analyzed to compare nAPCsr and nAPCsr10. These values were obtained using the validated ETP‐based APCr assay. Results The Spearman correlation between nAPCsr and nAPCsr10 had a coefficient of 0.99. Linear regression showed the following equation y = 0.9315*x + 0.03942 (r2 = .97). When differences (nAPCsr10 – nAPCsr) were plotted against nAPCsr10, the mean difference equaled 0.16% or 4.95%. The correction obtained with the use of the nAPCsr10 showed that the results of the nAPCsr were statistically different (P < .0001). Conclusions This new scale provides a harmonization and normalization of the nAPCsr. Results show a better reproducibility with the nAPCsr10. It avoids the additional variability and the unharmonized scale introduced by the use of a reference plasma. This adapted method for the calculation of the APC resistance could provide the regulatory and scientific bodies with more reproducible and harmonized evaluations.

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Section: Discussionsupporting

confidence: 57%

Section: Reference Plasma: Which One To Use and To Normalize What?supporting

confidence: 51%

Section: Reference Plasma: Which One To Use and To Normalize What?supporting

confidence: 51%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Proof of concept of a new scale for the harmonization and the standardization of the ETP‐based APC resistance

Morimont

Bouvy²,

Delvigne³

et al. 2020

Journal of Thrombosis and Haemostasis

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“…The use of standardized reagents and a highly automated process integrating different steps of TG measurement, including PRP separation, platelet count adjustment, and dispensing of plasma and reagents would highly contribute to uniformity of results across different laboratory settings and clinical conditions. A recently developed, fully automated TG analyzer (ST-Genesia, Stago, Asnières-sur-Seine, France), when tested using PFP with strict control of temperature, volumes, and normalization of each parameter against a reference plasma, offered increased reproducibility of the TG measurements [62,63]. Further investigations are needed to determine if the same principle could be applied to platelet-dependent TG.…”

Section: Uncertainties and Limitations Of Thrombin Generation In The mentioning

confidence: 99%

Clinical Applications, Pitfalls, and Uncertainties of Thrombin Generation in the Presence of Platelets

Panova‐Noeva

Meijden

Cate

2019

JCM

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Platelet-dependent thrombin generation is a helpful tool to assess ex vivo the interaction between platelets and plasma coagulation factors in the initiation, amplification, and inhibition of thrombin generation (TG). This review article discusses the most relevant available data on the clinical applications of fluorogenic TG, the most widely used TG assay, performed in the presence of platelets, i.e., in platelet-rich plasma. With respect to prothrombotic states, arterial hypertension and obesity were the most prominent cardiovascular conditions linked to increased platelet-dependent TG. In addition, platelet-associated hypercoagulability, assessed by the TG assay, has been shown in individuals with active cancer. In terms of bleeding, platelet-dependent TG has been applied to assess bleeding risk in individuals with hemophilia, von Willebrand disease, and Glanzmann thrombasthenia as well as in subjects with other congenital or acquired coagulation factor deficiencies. In addition to risk prediction, a role of the TG assay has been suggested in monitoring antiplatelet therapy in prothrombotic conditions and replacement therapy in bleeding diathesis. Finally, for the routine clinical use and as a biomarker of disease development and progression, better standardization and clinical validation of platelet-dependent TG are still needed.readings from a fluorometer are recorded and calculated by means of the Thrombinoscope software (Thrombinoscope BV). In principle, the measurements in PRP are assessed in fresh material, whereas TG in PFP is assessed in frozen, stored material. For TG measurements in PRP, coagulation is triggered by adding 20 µL exogenous tissue factor (TF) to 80 µL PRP (with adjusted platelet concentration of 150,000 platelets/µL), whereas for TG measurements in PFP, 20 µL exogenous TF and 4 µM phospholipids are added to 80 µL plasma. After prewarming for 10 min at 37 • C in the fluorimeter, the reaction is started by adding 20 µL of a low-affinity fluorogenic substrate for thrombin (Z-Gly-Gly-Arg-AMC) and calcium chloride mixture (FluCa). TG is measured as a function of a calibration signal obtained in a sample from the same plasma (PRP or PFP) after addition of a fixed amount of thrombin-α2-macroglobulin complex (20 µL of thrombin calibrator) and 20 µL of FluCa. The TG method is extensively reviewed in [5][6][7]. Parameters derived from a TG curve and most frequently reported in the studies are lag time, time to minimum thrombin formed expressed in minutes (min); peak height, maximum concentration of thrombin formed expressed in nM thrombin; and endogenous thrombin potential (ETP) or area under the curve expressed as nM of thrombin formed per minute.The aim of this article was to review the available evidence for the potential clinical application of platelet-dependent TG in both prothrombotic and bleeding conditions, as depicted in Figure 1. Aspects preventing its broader application and routine clinical practice will also be discussed as uncertainties and limitations of thrombin generation in the...

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Inhibition of thrombin generation 12 hours after intake of direct oral anticoagulants

Metze

Pfrepper

Klöter

et al. 2020

Research and Practice in Thrombosis and Haemostasis

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Background The residual antithrombotic activity 12 hours after intake of direct oral anticoagulants (DOACs) is of clinical relevance in the setting of bleeding or urgent surgery. Objective To evaluate the effects of DOACs on thrombin generation 12 hours after DOAC intake in comparison to baseline and a healthy control group. Methods Eighty patients were recruited, 20 patients for each approved DOAC: apixaban, edoxaban, rivaroxaban, and dabigatran. The patients were either to be put on anticoagulation for the first time or had stopped taking oral anticoagulation for at least 48 hours. Blood plasma was sampled before (baseline) and 12 hours after starting DOAC for quantification of drug levels and thrombin generation assayed using an automated system (ST Genesia). Sixty‐one blood donors served as control group. Results The factor Xa inhibitors significantly increased lag time (137%‐219%) and reduced thrombin peak (47%‐76%) and velocity index (17%‐44%) after 12 hours compared to baseline. Dabigatran showed prolongation of lag time to 133% and time to peak to 119%. All patients had residual antithrombotic activity, with reduced thrombin generation parameters 12 hours after DOAC intake compared to baseline and to the healthy control group. This effect remained significant in patients with low residual DOAC plasma levels <50 ng/mL. Conclusion Thrombin generation remains reduced 12 hours after DOAC intake. While thrombin peak is particularly modified by factor Xa inhibitors, all DOACs prolong the lag time and time to thrombin peak. In the setting of bleeding or urgent surgery, the automated thrombin generation assay may assist in decision making and antidote administration.

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Assessment of the analytical performances and sample stability on ST Genesia system using the STG‐DrugScreen application

Cited by 44 publications

References 36 publications

Proof of concept of a new scale for the harmonization and the standardization of the ETP‐based APC resistance

Proof of concept of a new scale for the harmonization and the standardization of the ETP‐based APC resistance

Clinical Applications, Pitfalls, and Uncertainties of Thrombin Generation in the Presence of Platelets

Inhibition of thrombin generation 12 hours after intake of direct oral anticoagulants

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