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Thrombin Generation Assays (TGAs)

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Hemostasis and Thrombosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1646))

Abstract

In recent years, there has been a revival of interest in a time-honored tool of the clotting trade: the thrombin generation curve or thrombin generation (TG) assay (TGA) as we now call it. Thrombin generation is a key coagulation process that determines the extent of a hemostatic plug or a thrombotic process. The recent development of newer tests based on the continuous registration of TG under in vitro conditions that mimic more closely what occurs in vivo prompted us to reinvestigate the balance between procoagulants and anticoagulants in patients. Measurement of TG is accepted as a research tool, but the variety of sources and concentrations of reagents, as well as technical constraints, limit its potential for proper clinical use. The newer TGAs not only provide an overall assessment of hemostasis but also provide promising laboratory tools for investigating hemorrhagic coagulopathies and monitoring replacement therapy in several clinical conditions.

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Abbreviations

AMC:

7-Amino-4-methylcoumarin

BSA:

Bovine serum albumin

CaCl2 :

Calcium chloride

DMSO:

Dimethyl sulfoxide

HEPES:

2-{−4-(2-Hydroxyethyl]-1-piperazinyl}-ethanesulfonic acid, HNa buffer, HEPES/NaCl buffer

HNaBSA5:

HEPES/NaCl/0.5% BSA buffer

Max slope:

maximum slope (RFU I min)

NaCl:

Sodium chloride

PC:

Phosphatidylcholine

PL:

Phospholipid

PS:

Phosphatidylserine

RFU:

Relative fluorescence unit(s)

RT:

Room temperature

rTF:

recombinant tissue factor

TF:

Tissue factor

Time max slope:

time of maximum slope velocity (sec)

Z-Gly-Gly-Arg-AMC.HCI:

Benzyloxylcarbonyl-glycine-glycine-arginine-7-amido-4-methylcoumarin hydrochloride.

References

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Acknowledgments

Maj Ekman and Margoth Gunnarsson are acknowledged for providing excellent technical assistance. Funding was obtained from Lund University and Region Skane, Sweden.

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Authors and Affiliations

  1. Section of Clinical Biochemistry, Department of Neurological, Biomedical and Movement Sciences, University of Verona, Piazzale L.A. Scuro 10, 37134, Verona, Italy

    Gian Luca Salvagno M.D., Ph.D.

  2. Center for Thrombosis and Hemostasis, Lund University, Skane University Hospital, Jan Waldenströms gata 14, SE-205 02, Malmö, Sweden

    Erik Berntorp

Authors
  1. Gian Luca Salvagno M.D., Ph.D.
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  2. Erik Berntorp
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Corresponding author

Correspondence to Gian Luca Salvagno M.D., Ph.D. .

Editor information

Editors and Affiliations

  1. Department of Haematology, Sydney Centres for Thrombosis and Haemostasis, Institute of Clinical Pathology and Medical Research (ICPMR), NSW Health Pathology, Westmead Hospital, Westmead, New South Wales, Australia

    Emmanuel J. Favaloro

  2. Section of Clinical Biochemistry, University of Verona, Verona, Italy

    Giuseppe Lippi

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Salvagno, G.L., Berntorp, E. (2017). Thrombin Generation Assays (TGAs). In: Favaloro, E., Lippi, G. (eds) Hemostasis and Thrombosis. Methods in Molecular Biology, vol 1646. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7196-1_37

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  • DOI: https://doi.org/10.1007/978-1-4939-7196-1_37

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7194-7

  • Online ISBN: 978-1-4939-7196-1

  • eBook Packages: Springer Protocols

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