Optimisation of a new continuous UV assay for the determination of blood coagulation factor XIII activity in human plasma.
The new photometric assay described by Fickenscher et al. (Thromb. Haemostas. 65 (1991) 535-540) for the determination of factor XIII facilitates the diagnosis of factor XIII deficiency. In spite of easy handling, this test should be used critically. Patients with hyperfibrinogenaemia showed factor XIII activities of less than 20%, whereas with an optimized assay we found normal factor XIII values. Also, the use of a fixed period of incubation for the analysis is questionable, because the period of constant reaction rate occurs earlier and is shorter with high factor XIII activities and later and longer with low factor XIII activities. A linear relation between factor XIII activity and signal only exists up to 80% of activity. In some plasma samples from patients with hyperfibrinogenaemia the factor XIII determination actually shows decreased values for factor XIII. During the reaction, a fibrin clot is formed. The resulting turbidity simulates an increase in absorbance so that NADH consumption is apparently decreased. In six patients with hyperfibrinogenaemia (8.1-9.4 g/l), a factor XIII activity of 26 U/l or less was determined. Using 50 microliters instead of 100 microliters sample volume, 50% (3/6) of the patients showed a normal factor XIII activities (80-96 U/l), whereas 50% (3/6) values of 6-15 U/l were found. In our modified assay we measured normal factor XIII activities (72-151 U/l) in all 6 patients. The procedure is optimized by reducing the sample volume from 100 microliters to 50 microliters.(ABSTRACT TRUNCATED AT 250 WORDS)
Heins
M
M
Institut für Klinische Chemie und Laboratoriumsdiagnostik, Medizinische Einrichtungen, Heinrich-Heine-Universität, Düsseldorf, Germany.
Fahron
U
U
Withold
W
W
Rick
W
W
D005176:Factor XIII / Q000032:analysis*; D005340:Fibrinogen / Q000032:analysis; D006801:Humans; D007700:Kinetics; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D005340:Fibrinogen; D005176:Factor XIII