Cargando…

Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability

INTRODUCTION: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection is associated with a clear prothrombotic phenotype. Although the exact pathophysiological mechanisms are not yet fully understood, thrombosis is clearly a highly important in the prognosis and outcome of COVID-19. As...

Descripción completa

Detalles Bibliográficos
Autores principales: van de Berg, Tom W., Hulshof, Anne-Marije M., Nagy, Magdolna, van Oerle, Rene, Sels, Jan-Willem, van Bussel, Bas, ten Cate, Hugo, Henskens, Yvonne, Spronk, Henri M.H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Author(s). Published by Elsevier Ltd. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7903905/
https://www.ncbi.nlm.nih.gov/pubmed/33662799
http://dx.doi.org/10.1016/j.thromres.2021.02.026
_version_ 1783654825209102336
author van de Berg, Tom W.
Hulshof, Anne-Marije M.
Nagy, Magdolna
van Oerle, Rene
Sels, Jan-Willem
van Bussel, Bas
ten Cate, Hugo
Henskens, Yvonne
Spronk, Henri M.H.
author_facet van de Berg, Tom W.
Hulshof, Anne-Marije M.
Nagy, Magdolna
van Oerle, Rene
Sels, Jan-Willem
van Bussel, Bas
ten Cate, Hugo
Henskens, Yvonne
Spronk, Henri M.H.
author_sort van de Berg, Tom W.
collection PubMed
description INTRODUCTION: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection is associated with a clear prothrombotic phenotype. Although the exact pathophysiological mechanisms are not yet fully understood, thrombosis is clearly a highly important in the prognosis and outcome of COVID-19. As such, there is a need for diagnostic analysis and quantification of the coagulation potential in these patients, both at diagnosis and follow-up. Global coagulation assays like thrombin generation (TG) and rotational thromboelastometry (ROTEM) might be suitable in estimating COVID-19 associated coagulopathy and thrombosis risk. Therefore, we aimed at validating both assays for samples with high levels of fibrinogen and in the presence of anticoagulant heparins, such as commonly observed for COVID-19 ICU patients. MATERIALS AND METHODS: Calibrated Automated Thrombography (CAT) was optimized to assess plasma thrombin generation in the presence of heparins. The final conditions with either 10 μg/mL Ellagic acid (EA) or PPP Reagent HIGH (high tissue factor; HPPH) were validated according to the EP5 protocol for within-run and between-run variability. Overall variability was well below 10%. To estimate the influences of heparins and high fibrinogen levels, CAT was performed on spiked plasma aliquots from 13 healthy volunteers. Comparable to the CAT method, tPA-ROTEM was used to validate the effect of high fibrinogen and heparins on clotting time, clot firmness and clot lysis parameters. RESULTS: Our adjusted COVID-19 assay showed a heparin dose dependent decrease in peak height and endogenous thrombin potential (ETP) for both EA and HPPH triggered variants. High fibrinogen did not alter the inhibitory effect of either LMWH or UFH, nor did it influence the peak height or ETP in any of the conditions. The tPA-ROTEM showed a significant prolongation in clotting time with the additions of heparin, which normalized with the addition of high fibrinogen. MCF was markedly increased in all hyperfibrinogenemic conditions. A trend towards increased lysis time and, thus, decreased fibrinolysis was observed. CONCLUSION: Thrombin generation and tPA-ROTEM protocols for measurements in the COVID-19 populations were adjusted and validated. The adjusted thrombin generation assay shows good sensitivity for measurements in heparin spiked plasma. High levels of fibrinogen did not alter the assay or the effectiveness of heparins as measured in this assay. t-PA ROTEM was effective in measurement of both high fibrinogen and heparins spiked samples and was sensitive to the expected relevant coagulant changes by these conditions. No clear fibrinolytic effect was observed in different conditions.
format Online
Article
Text
id pubmed-7903905
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher The Author(s). Published by Elsevier Ltd.
record_format MEDLINE/PubMed
spelling pubmed-79039052021-02-25 Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability van de Berg, Tom W. Hulshof, Anne-Marije M. Nagy, Magdolna van Oerle, Rene Sels, Jan-Willem van Bussel, Bas ten Cate, Hugo Henskens, Yvonne Spronk, Henri M.H. Thromb Res Full Length Article INTRODUCTION: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection is associated with a clear prothrombotic phenotype. Although the exact pathophysiological mechanisms are not yet fully understood, thrombosis is clearly a highly important in the prognosis and outcome of COVID-19. As such, there is a need for diagnostic analysis and quantification of the coagulation potential in these patients, both at diagnosis and follow-up. Global coagulation assays like thrombin generation (TG) and rotational thromboelastometry (ROTEM) might be suitable in estimating COVID-19 associated coagulopathy and thrombosis risk. Therefore, we aimed at validating both assays for samples with high levels of fibrinogen and in the presence of anticoagulant heparins, such as commonly observed for COVID-19 ICU patients. MATERIALS AND METHODS: Calibrated Automated Thrombography (CAT) was optimized to assess plasma thrombin generation in the presence of heparins. The final conditions with either 10 μg/mL Ellagic acid (EA) or PPP Reagent HIGH (high tissue factor; HPPH) were validated according to the EP5 protocol for within-run and between-run variability. Overall variability was well below 10%. To estimate the influences of heparins and high fibrinogen levels, CAT was performed on spiked plasma aliquots from 13 healthy volunteers. Comparable to the CAT method, tPA-ROTEM was used to validate the effect of high fibrinogen and heparins on clotting time, clot firmness and clot lysis parameters. RESULTS: Our adjusted COVID-19 assay showed a heparin dose dependent decrease in peak height and endogenous thrombin potential (ETP) for both EA and HPPH triggered variants. High fibrinogen did not alter the inhibitory effect of either LMWH or UFH, nor did it influence the peak height or ETP in any of the conditions. The tPA-ROTEM showed a significant prolongation in clotting time with the additions of heparin, which normalized with the addition of high fibrinogen. MCF was markedly increased in all hyperfibrinogenemic conditions. A trend towards increased lysis time and, thus, decreased fibrinolysis was observed. CONCLUSION: Thrombin generation and tPA-ROTEM protocols for measurements in the COVID-19 populations were adjusted and validated. The adjusted thrombin generation assay shows good sensitivity for measurements in heparin spiked plasma. High levels of fibrinogen did not alter the assay or the effectiveness of heparins as measured in this assay. t-PA ROTEM was effective in measurement of both high fibrinogen and heparins spiked samples and was sensitive to the expected relevant coagulant changes by these conditions. No clear fibrinolytic effect was observed in different conditions. The Author(s). Published by Elsevier Ltd. 2021-05 2021-02-24 /pmc/articles/PMC7903905/ /pubmed/33662799 http://dx.doi.org/10.1016/j.thromres.2021.02.026 Text en © 2021 The Author(s) Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Full Length Article
van de Berg, Tom W.
Hulshof, Anne-Marije M.
Nagy, Magdolna
van Oerle, Rene
Sels, Jan-Willem
van Bussel, Bas
ten Cate, Hugo
Henskens, Yvonne
Spronk, Henri M.H.
Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability
title Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability
title_full Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability
title_fullStr Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability
title_full_unstemmed Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability
title_short Suggestions for global coagulation assays for the assessment of COVID-19 associated hypercoagulability
title_sort suggestions for global coagulation assays for the assessment of covid-19 associated hypercoagulability
topic Full Length Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7903905/
https://www.ncbi.nlm.nih.gov/pubmed/33662799
http://dx.doi.org/10.1016/j.thromres.2021.02.026
work_keys_str_mv AT vandebergtomw suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT hulshofannemarijem suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT nagymagdolna suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT vanoerlerene suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT selsjanwillem suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT vanbusselbas suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT tencatehugo suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT henskensyvonne suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT spronkhenrimh suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability
AT suggestionsforglobalcoagulationassaysfortheassessmentofcovid19associatedhypercoagulability