n Medical Technology SA - A rapid and cost-effective method to visualize von Willebrand factor multimers in plasma
|Article Title||A rapid and cost-effective method to visualize von Willebrand factor multimers in plasma|
|© Publisher:||The Society of Medical Laboratory Technologists of South Africa (SMLTSA)|
|Journal||Medical Technology SA|
|Author||Muriel Meiring, Philip N. Badenhorst and Mareli Kelderman|
|Publication Date||Dec 2005|
|Pages||16 - 18|
The multimeric analysis of von Willebrand factor (VWF) is utilised for von Willebrand factor disease (VWD) subclass identification and is important for treating purposes. A highly sensitive and rapid method for the visualisation of the multimeric structure of VWF in plasma and platelets was described by Krizek et al in 2000 . This method uses a western blot technique where horizontal agarose electrophoresis is followed by the transfer of the VWF onto a polyvinylidine fluoride (PVDF) membrane. The multimeric pattern of VWF is visualized by immunolocalisation and luminographic detection and no radioactivity is used. We modified this method comprehensively to increase its sensitivity and to reduce the cost and duration of the test. We used one in stead of two localisation antibodies and thereby reduced the immunolocalisation time by more than two hours. This also reduced the cost of the procedure. We further reduced the cost by using two carbon plates for blotting in stead of a blotter instrument. A total cost reduction of 40% could be achieved. A higher sensitivity was obtained by degassing the agarose before the casting process. The higher sensitivity is reflected by the fact that differences between the multimer patterns of type 2M and normal patients could be detected.
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