Abstract Number: PB0603
Meeting: ISTH 2020 Congress
Background: Haemolysis, i.e. the release of intracellular components of blood cells, is visible as a reddish colorization of the plasma after centrifugation. This phenomenon should not be ignored as it can interfere with laboratory measurements both because of optical disturbances (released haemoglobin absorbs light at specific wavelengths) and coagulation activation through membrane components exposure. Distinguishing both contributors can be of help in understanding observation of an impact of haemolysis on an assay.
Aims: To investigate whether the presence of haemoglobin in plasma samples can interfere with the measurement of thrombin generation (TG) with ST Genesia.
Methods: Plasma samples (N=3) were prepared by mixing a fixed volume of plasma (475 µl) with a fixed volume of buffer (25 µl) containing a varying amount of haemoglobin. In total, 7 samples were prepared with a haemoglobin concentration ranging from 0 mg/dl up to 190 mg/dl. An in-house prothrombin time assay on the STart was used to exclude the presence of heparin in the hemoglobin preparation and TG was measured with the ST Genesia using STG-BleedScreen reagent.
Results: The in-house prothrombin measurements revealed no presence of heparin in the samples. Normalized results (mean (±SD)) of lagtime, time-to-peak, peak height and endogenous thrombin potential were 99.6% (±3.7%), 101.0% (±3.7%), 99.5% (±6.2%) and 98.2% (±3.5), respectively. One-way ANOVA multiple comparison revealed no significant differences between the baseline sample without haemoglobin and all other samples containing haemoglobin (p>0.05).
Conclusions: No differences in TG could be observed in samples containing varying haemoglobin concentrations. The calibration method of the ST Genesia is able to correct for the colour of the plasma samples in the presence of haemoglobin. Any effect of haemolysis on TG would then be due to activation of coagulation with cell components and not to the colour of the released haemoglobin.
To cite this abstract in AMA style:Ninivaggi M, Yan Q, de Laat-Kremers R, Carlo A, de Laat B. Measuring Thrombin Generation in the Presence of Haemoglobin on the ST Genesia [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/measuring-thrombin-generation-in-the-presence-of-haemoglobin-on-the-st-genesia/. Accessed December 10, 2023.
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