Large external quality assessment survey on thrombin generation with CAT: further evidence for the usefulness of normalisation with an external reference plasma

Perrin, Julien; Depasse, François; Lecompte, Thomas; Raucourt, Emmanuelle de; Planche, Vincent; Ajzenberg, Nadine; Ollivier, Véronique; Helley, Dominique; Dignat-George, Françoise; Morange, P.-E.; Drouet, Ludovic; Samama, M; Minon, Jean-Marc; Freyburger, G.; Mingant, Fanny; Sinegre, Thomas; Dutrillaux, F.; Marlu, Raphaël; Jeanpierre, Emmanuelle; Dargaud, Yesim; Sauguet, Pauline; Gris, Jean‐Christophe; Duchemin, Jérôme; Hézard, Nathalie; Guéret, P.; Cam-Duchez, Véronique Le; Tardy, B.; Delahousse, B.; Ravanat, Catherine; Hanss, Michel; Fontana, Pierre; Feugeas, Olivier; Gheldof, Damien; Douxfils, Jonathan

doi:10.1016/j.thromres.2014.12.015

Cited by 61 publications

(81 citation statements)

References 27 publications

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“…It was not our aim in this study to develop a reference range tested on healthy individuals and therefore no such population was enrolled here. Although standardization of the TGT is far from being completed, optimal experimental conditions have been described previously, which were followed by us in this study [26, 27]. This includes, among others, the use of a standardized thrombin calibrator, which allows the quantitative expression of thrombin, the use of the Thrombogram software, which can reliably describe the distinct phases of thrombin generation, and the practice that all samples were run in duplicates.…”

Section: Discussionmentioning

confidence: 99%

“…Thrombin generation test was performed as described previously using the Thrombinoscope CAT (Calibrated Automated Thrombogram, Maastricht, The Netherlands) assay according to the manufacturer’s instructions (Diagnostica Stago, Asnières, France) [7,8]. Briefly, 80 microliters of plasma was incubated with 20 μL PPP-Reagent ™ (containing 5 pM recombinant tissue factor and 4 μM phospholipids) for 10 minutes in round-bottomed 96-well black microplates.…”

Section: Methodsmentioning

confidence: 99%

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Low thrombin generation predicts poor prognosis in ischemic stroke patients after thrombolysis

et al. 2017

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Thrombolysis by intravenous recombinant tissue plasminogen activator (rt-PA) is an effective therapy in acute ischemic stroke (AIS). Thrombin generation test (TGT) is a global hemostasis test providing information about the speed and amount of generated thrombin in plasma. Here we aimed to find out whether results of this test before the initiation of thrombolysis might predict outcomes. Study population included 120 consecutive AIS patients, all within 4.5 hours of their symptom onset, who underwent thrombolysis by rt-PA. Blood samples were collected from all patients upon admission and TGT was performed using platelet poor plasma. Clinical data of patients including the NIHSS were registered at admission, day 1 and 7 after therapy. The ASPECT score was assessed using CT images taken before and 24 hours after thrombolysis. Long-term functional outcome was defined 3 months after the event by the modified Rankin Scale. Endogenous Thrombin Potential (ETP) and Peak Thrombin were significantly lower in patients with cardioembolic IS. Symptomatic intracranial hemorrhage (SICH) was found in 6 patients and was significantly associated with low ETP and Peak Thrombin levels. A multiple logistic regression model revealed that an ETP result in the lower quartile is an independent predictor of mortality within the first two weeks (OR: 6.03; 95%CI: 1.2–30.16, p<0.05) and three months after the event (OR: 5.28; 95%CI: 1.27–21.86, p<0.05). Low levels of ETP and Peak Thrombin parameters increase the risk of therapy associated SICH. A low ETP result is an independent predictor of short- and long-term mortality following thrombolysis.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Low thrombin generation predicts poor prognosis in ischemic stroke patients after thrombolysis

et al. 2017

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“…has shown that by standardizing protocols for the Thrombinoscope, through training along with use of standard materials, could help transition thrombin generation from the laboratory to general clinical practice 7. Further research shows that using normalized, standard reference plasma samples between multiple centers along with strict adherence to protocols is critical to obtain clinically valid results 32, 33…”

Section: Thrombin Generation In Clincal Setttingsmentioning

confidence: 99%

A review of commercially available thrombin generation assays

Kintigh

Monagle

Ignjatović

2018

Research and Practice in Thrombosis and Haemostasis

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Essentials Overview of the three commercial thrombin generation methods.Description of the sample preparation, data management, and analysis.Description of the similarities and differences in regards to substrates results.Discussion of advantages and disadvantages of the three approaches. Currently there are three commercially available thrombin generation methods. These methods help detect the levels of thrombin generated in patient samples by the use of chromogenic or fluorogenic substrates in plasma or whole blood. Determining the rate of thrombin generation can help indicate if patients are at risk of clotting or bleeding. This review discusses two fluorogenic and one chromogenic method and focuses on similarities and differences of these three methods. The review specifically focuses on the accuracy of commercial substrates used in thrombin generation, and interference that can occur by various plasma proteins, as well as on evaluating the advantages and disadvantages of each method. The commercial chromogenic assay and both fluorogenic assays are able to monitor the rate of thrombin generation and can give indications towards potential coagulation abnormalities. Overall, the main differences between the thrombin generation methods are based on the type of substrate used, sample preparation, and data processing. Despite advancement in this field there are still technical challenges that preclude the widespread use of thrombin generation in clinical applications.

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“…Furthermore, it has the ability to be available in much larger standardised batches than normal pooled plasma and can be used countrywide or even worldwide. Indeed, a recent study has indicated that normal pooled plasma produced locally does not provide as standardised results as that produced externally [22]. Finally, it has been shown that lyophilised plasmas produce the same shaped thrombin generation curve as that obtained from normal pooled plasma, suggesting that it provides relevant results [9].…”

Section: Limitationsmentioning

confidence: 99%

Establishing a reference range for thrombin generation using a standard plasma significantly improves assay precision

Bagot

Leishman

2015

Thrombosis Research

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Large external quality assessment survey on thrombin generation with CAT: further evidence for the usefulness of normalisation with an external reference plasma

Cited by 61 publications

References 27 publications

Low thrombin generation predicts poor prognosis in ischemic stroke patients after thrombolysis

Low thrombin generation predicts poor prognosis in ischemic stroke patients after thrombolysis

A review of commercially available thrombin generation assays

Establishing a reference range for thrombin generation using a standard plasma significantly improves assay precision

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