Cargando…

Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay

The goat model of atrial fibrillation (AF) allows investigation of the effect of AF on coagulation. However, assays for goat plasma are not available from commercial sources. Calibrated automated thrombography (CAT) provides a global view of the coagulation profile by assessing in vitro thrombin gen...

Descripción completa

Detalles Bibliográficos
Autores principales: D'Alessandro, Elisa, Scaf, Billy, van Oerle, René, van Nieuwenhoven, Frans A., van Hunnik, Arne, Verheule, Sander, Schotten, Ulrich, ten Cate, Hugo, Spronk, Henri M. H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8606030/
https://www.ncbi.nlm.nih.gov/pubmed/34849448
http://dx.doi.org/10.1002/rth2.12620
_version_ 1784602268698411008
author D'Alessandro, Elisa
Scaf, Billy
van Oerle, René
van Nieuwenhoven, Frans A.
van Hunnik, Arne
Verheule, Sander
Schotten, Ulrich
ten Cate, Hugo
Spronk, Henri M. H.
author_facet D'Alessandro, Elisa
Scaf, Billy
van Oerle, René
van Nieuwenhoven, Frans A.
van Hunnik, Arne
Verheule, Sander
Schotten, Ulrich
ten Cate, Hugo
Spronk, Henri M. H.
author_sort D'Alessandro, Elisa
collection PubMed
description The goat model of atrial fibrillation (AF) allows investigation of the effect of AF on coagulation. However, assays for goat plasma are not available from commercial sources. Calibrated automated thrombography (CAT) provides a global view of the coagulation profile by assessing in vitro thrombin generation (TG). We describe the customization of the CAT assay in goat platelet‐poor plasma (PPP) and in factor Xa (FXa)‐inhibitor‐anticoagulated PPP. TG was initiated in the presence of phospholipids and either (a) PPP reagent, reagent low, or reagent high; (b) goat brain protein extraction (GBP); or (c) Russell's viper venom‐factor X activator (RVV‐X). Contact activation was assessed by adding corn trypsin inhibitor. Different concentrations of prothrombin complex concentrate (PCC) were used to determine the sensitivity of both the GBP and RVV‐X method. To obtain FXa‐inhibitor anticoagulated plasma, rivaroxaban was added to plasma. TG settings with human reagents were not suitable for goat plasma. TG triggered with GBP increased peak height and ETP values. Similarly, the RVV‐X method produced comparable TG curves and was more sensitive to PCC titration. Finally, both methods were able to detect the decrease in clotting potential induced by FXa inhibition. This is the first study that reports the customization of the CAT assay for goats. The GBP and RVV‐X methods were comparable in triggering TG in goat plasma. The RVV‐X method seemed to better discriminate changes in TG curves due to increases in clotting potential as well as to FXa inhibition by rivaroxaban in goat plasma.
format Online
Article
Text
id pubmed-8606030
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-86060302021-11-29 Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay D'Alessandro, Elisa Scaf, Billy van Oerle, René van Nieuwenhoven, Frans A. van Hunnik, Arne Verheule, Sander Schotten, Ulrich ten Cate, Hugo Spronk, Henri M. H. Res Pract Thromb Haemost Methodological Article The goat model of atrial fibrillation (AF) allows investigation of the effect of AF on coagulation. However, assays for goat plasma are not available from commercial sources. Calibrated automated thrombography (CAT) provides a global view of the coagulation profile by assessing in vitro thrombin generation (TG). We describe the customization of the CAT assay in goat platelet‐poor plasma (PPP) and in factor Xa (FXa)‐inhibitor‐anticoagulated PPP. TG was initiated in the presence of phospholipids and either (a) PPP reagent, reagent low, or reagent high; (b) goat brain protein extraction (GBP); or (c) Russell's viper venom‐factor X activator (RVV‐X). Contact activation was assessed by adding corn trypsin inhibitor. Different concentrations of prothrombin complex concentrate (PCC) were used to determine the sensitivity of both the GBP and RVV‐X method. To obtain FXa‐inhibitor anticoagulated plasma, rivaroxaban was added to plasma. TG settings with human reagents were not suitable for goat plasma. TG triggered with GBP increased peak height and ETP values. Similarly, the RVV‐X method produced comparable TG curves and was more sensitive to PCC titration. Finally, both methods were able to detect the decrease in clotting potential induced by FXa inhibition. This is the first study that reports the customization of the CAT assay for goats. The GBP and RVV‐X methods were comparable in triggering TG in goat plasma. The RVV‐X method seemed to better discriminate changes in TG curves due to increases in clotting potential as well as to FXa inhibition by rivaroxaban in goat plasma. John Wiley and Sons Inc. 2021-11-21 /pmc/articles/PMC8606030/ /pubmed/34849448 http://dx.doi.org/10.1002/rth2.12620 Text en © 2021 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis (ISTH). https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Methodological Article
D'Alessandro, Elisa
Scaf, Billy
van Oerle, René
van Nieuwenhoven, Frans A.
van Hunnik, Arne
Verheule, Sander
Schotten, Ulrich
ten Cate, Hugo
Spronk, Henri M. H.
Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay
title Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay
title_full Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay
title_fullStr Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay
title_full_unstemmed Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay
title_short Thrombin generation by calibrated automated thrombography in goat plasma: Optimization of an assay
title_sort thrombin generation by calibrated automated thrombography in goat plasma: optimization of an assay
topic Methodological Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8606030/
https://www.ncbi.nlm.nih.gov/pubmed/34849448
http://dx.doi.org/10.1002/rth2.12620
work_keys_str_mv AT dalessandroelisa thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT scafbilly thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT vanoerlerene thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT vannieuwenhovenfransa thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT vanhunnikarne thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT verheulesander thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT schottenulrich thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT tencatehugo thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay
AT spronkhenrimh thrombingenerationbycalibratedautomatedthrombographyingoatplasmaoptimizationofanassay