Von Willebrand factor multimeric assay: novel diagnostics capabilities

Author:

Poletaev A. V.1ORCID,Seregina E. A.2ORCID,Pshonkin A. V.1ORCID,Karamyan N. A.1ORCID,Fedorova D. V.1ORCID,Plyasunova S. A.1ORCID,Zharkov P. A.1ORCID

Affiliation:

1. Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Ministry of Health of Russia

2. Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Ministry of Health of Russia; Center for Theoretical Problems of Physicochemical Pharmacology of the Russian Academy of Sciences

Abstract

Introduction. Distinguishing between Von Willebrand disease (vWD) types often requires multimer gel analysis. The current techniques for vWF multimer structure are manual, complicated, non-standardized and time consuming. The aim of this study was to evaluate diagnostic capabilities of new automated vWF multimer screening assay.Materials and methods. Children with vWD, acquired von Willebrand Syndrome (aVWS) and 8 healthy donors as a control group were enrolled in this study. Von Willebrand factor antigen (vWF Ag); ristocetin cofactor activity (VWF:Rco); vWF collagen binding (VWF:CB); ristocetin-induced platelet aggregation (RIPA); factor VIII clotting activity (FVIII:C) and vWF factor VIII binding activity (vWF:FVIIIb) were performed to evaluate vWD. Multimer analysis was carried out using the commercial HYDRAGEL 5 von Willebrand Multimers kit on semi-automatic gel electrophoresis instrument HYDRASYS (SEBIA).Results. The samples from control group had 9—12 bands of vWF multimers with the same distribution as control plasma. Patients with type I vWD had the proportional decrease in the intensity of the bands with preservation of the normal distribution of the band. Patients with type III vWD reveal the complete absence of the multimer bands on the gel. Multimer analysis in type IIA shows the absence of high molecular weight multimer bands. In other patients the distribution of vWF multimers was normal against the changes in functional properties of vWF (types IIM, N). Most of the children with aVWS also revealed normal distribution of vWF multimers, however, in some patients, the slight decrease in large multimeric forms was observed visually on the gel.Conclusion. Multimer analysis allows to visualize the multimer distribution in various types of von Willebrand disease. The method is easy to perform and can be useful for distinguishing between the subtypes of vWD. But only the full test panel including genetic tests would allow the differentiantion of vWD types with high precision.

Publisher

OOO Grafika

Subject

Oncology,Hematology,Pediatrics, Perinatology and Child Health

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