Latex Agglutination Method for Determination of von Willebrand Factor Antigen in Patients with Bleeding Disorders

Abstract

Measurement of the level of von Willebrand factor antigen (VWF:Ag) in human plasma is usually carried out in a specialized laboratory in routine clinical practice and is carried out by the ELISA method which takes a long time or immunoturbidimetry which is less sensitive and specific. The purpose of this study was to qualitatively determine the von Willebrand factor (VWF) antigen in only a maximum of 5 min using the latex agglutination method. The research design is an exploratory laboratory experiment. This study was conducted in a cross sectional manner in patients with bleeding disorders with prolonged bleeding time and healthy individuals. The research stage is to prepare a VWF polyclonal antibody from Invitrogen Ref. Pa5-16634 with Lot No. XE3587846A in 0.1M MES{2-(N-morpholino) ethane-sulfonic acid buffer, 0.9% sodium chloride, pH 6, to be sensitized to Carboxyl latex, 4% w/v 1 m. Latex particles after passive adsorption, polyclonal antibodies stick to the latex surface. The patient's citrate blood sample is taken which will be compared with the normal individual's citrate blood. The results showed agglutination in patients with bleeding disorders and no agglutination in normal individuals. In conclusion, latex reagent containing polyclonal VWF can be used as a diagnostic tool to determine VWF antigen in patients with bleeding disorders.