Abstract Number: PB0933
Meeting: ISTH 2021 Congress
Background: Multimeric assay is an important diagnostic tool in typing von Willebrand disease. However, this method allows only a visual assessment of the multimeric profile.
Aims: To develop an algorithm for the quantitative evaluation of von Willebrand factor multimers.
Methods: The semi-automatic method for evaluating multimers HYDRAGEL 5 vov Willebrand Multimers (Sebia, France) was used. The molecular ladder 1 Kb DNA Ladder M12 10000bp and Thermo Scientific FastRuler DNA Radder 100-5000 were used to assess the throughput of the gel, followed by staining with ethidium bromide. The gel of patients with established diagnosis of vWD and aVWS was evaluated, and PNP was used as a normal control. For a quantitative assessment of the photograph of the gel, the Imagej program was used, where the brightness of the bands was estimated by fractions of HF multimers – low(LMWM), medium(IMWM), high molecular weight(HMWM), and total amount.
Results: On gel with molecular ladder, 8 heavy and 2 medium ladder marks were visualized. The 8000bp label according to a molecular weight of 5200 kDa, which allows the separation of high and medium molecular weight forms, the border runs between 3 and 4 bands on the gel. The 4000bp label according to 2600 kDa, separating the medium and low molecular forms between bands 1 and 2. Accordingly, the first band on gel – LMWM, 2,3 bands – IMWM, 4 and further – HMWM. There was a good correlation(Pearson) between vWF:Ag and the overall brightness of the bands for all patients-0.95(p<0,01). Correlation between the brightness of HMWM and vWF:Rco in patients with vWD-0.99(p<0,01), in patients with aVWS – 0.54(p<0,34).
Conclusions: Evaluation of throughput capacity of the gel allowed to define the limits of the various forms multimers, which in fact differ from published data. The quantification algorithm has shown excellent correlations with vWF:Rco and vWF:Ag, and needs more research.
To cite this abstract in AMA style:Poletaev A, Seregina E, Karamyan N, Zharkov P. A Quantitative Evaluation of vWF Multimeric Structure in Patients with Different Types of vWD and aVWS [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/a-quantitative-evaluation-of-vwf-multimeric-structure-in-patients-with-different-types-of-vwd-and-avws/. Accessed December 7, 2021.
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