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Quantification of coagulation factor XIII activity by a thio-NADH based assay using factor XIII immuno-depleted plasma as a diluent for calibration

  • Andreas Kappel , Sina Stephan , Gerlinde Christ , Lena Pechmann , Christa Duwe , Bodo Fischer , Martin Hahn , Harald Althaus , Matthias Ehm , Herbert Schwarz and Frank Vitzthum
Published/Copyright: September 10, 2010

Abstract

Background: Accurate determination of factor XIII (FXIII) activity is crucial for replacement therapy. FXIII activity is typically determined using a coupled enzymatic reaction that measures nicotinamide adenine dinucleotide hydride (NADH) consumption at 340 nm.

Methods: Here, we describe the development of a prototype for a novel FXIII activity assay for detection at 405 nm by replacing NADH with thio-NADH, and the application of FXIII immuno-depleted plasma as a diluent for calibration.

Results: Performance data show up to two-fold lower susceptibility of the prototype assay to interferences from hemolyzed, icteric, and lipemic samples when compared to a NADH assay format. In addition, the use of FXIII immuno-depleted plasma as diluent for calibration improved recovery almost two-fold in the lower measurement range. The novel prototype assay correlates well with a conventional assay (r=0.98, y=0.99·x+2.17% FXIII, n=173).

Conclusions: The described prototype assay has the potential to (a) increase trueness of measurement of low levels of FXIII, (b) improve robustness due to reduction from interferences, and (c) can be used on a broad range of coagulation instruments due to its detection at 405 nm.

Clin Chem Lab Med 2010;48:1739–43.


Corresponding author: Frank Vitzthum, Siemens Healthcare Diagnostics Products GmbH, Pre-Development, P.O. Box 1149, 35001 Marburg, Germany Fax: +49 6421 39 3144,

Received: 2010-4-1
Accepted: 2010-6-17
Published Online: 2010-09-10
Published in Print: 2010-12-01

©2010 by Walter de Gruyter Berlin New York

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