Abstract Number: PB1130
Meeting: ISTH 2020 Congress
Background: Conventional coagulation assays are associated with certain limitations, as they lack to depict the clinical heterogeneity of bleeding in hemophilic patients. Global hemostasis assays (GHA) like thromboelastometry (ROTEM), thrombin generation assays (TGA) or clot waveform analysis (CWA) are helpful which can mimic and reflect the major physiological aspects of hemostasis process. However, these methods are still not well standardised and do not reflect correctly to the individual patient response to treatment with bypassing agents or novel factor concentrates. So, there is a need to make the GHAs like ROTEM and CWA more sensitive to correlate with the gold standard assay i.e. Thrombin generation assays.
Aims: To standardize and increase the sensitivity of ROTEM and CWA by using different combinations of phospholipid and tissue factor reagents.
Methods: Plasma samples of different factor levels (< 1IU/ml, 1IU/ml, 5IU/ml, 30IU/ml, 35IU/ml, 40IU/ml, 100IU/ml) were prepared by using different dilutions of factor concentrate in FVIII deficient plasma to mimic samples with different ETP values. The parameters used were ETP, Peak Height (ST-Genesia, Stago); 2nd Derivative +: CWA (ACL-TOP CTS 300, IL): ROTEM (ROTEM® delta Thromboelastometry System). The TGA was done in fully automated TGA (ST-Genesia, Stago). Phospholipid vesicles consisted of 20mol% phosphatidylserine, 20mol% phosphatidylethanolamine and 60mol% phosphatidylcholine and recombinant relipidated tissue factor not containing polybrene or Ca2+ from Dade Behring.
Results: In CWA, 2nd Derivative (+) shows statistically significant correlation with ETP when 1:2 Actin FS (Siemens Healthcare Diagnostics) was used as the triggering reagent (P< 0.05). ROTEM showed statistically significant correlation with ETP in both Whole blood and Platelet poor plasma (PPP) by using 0.5pM TF+2µM phospholipid and 0.2pM TF+0.8µM phospholipid as triggering reagent respectively (P< 0.05) [Table 1].
Conclusions: In the current study, GHAs like CWA and ROTEM were made sensitive to ETP with a combination of PL and TF reagents.
|FVIII levels (IU/ml)|
|GHA||Parameter||Difference between different FVIII levels: (Median)|
|CWA||2nd Derivative +||141||49||93||117.7|
|ROTEM||Clotting Time (CT)||88.5||50||103.5||141|
|TGA Genesia (Gold Std)||Endogenous Thrombin Potential (ETP)||69.9||69.16||139.09||114.83|
[Table 1: Difference in varying level of FVIII (IU/ml) and correlation with different GHAs.]
To cite this abstract in AMA style:Soni P, Patil R, Shetty S. Improved Sensitivity of Global Hemostasis Assays Using Different Combinations of Phospholipid Reagent and Tissue Factor [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/improved-sensitivity-of-global-hemostasis-assays-using-different-combinations-of-phospholipid-reagent-and-tissue-factor/. Accessed January 28, 2022.
« Back to ISTH 2020 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/improved-sensitivity-of-global-hemostasis-assays-using-different-combinations-of-phospholipid-reagent-and-tissue-factor/