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Updated: Aug 19, 2026

Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay
Published on: September 9, 2012
[Comparison of factor VIII:C and factor IX sensitivity of different commercial APTT reagents for canine plasma]
1Klinik für kleine Haustiere, Tierärztlichen Hochschule Hannover.
Abstract:
In the present study, six commercial reagents for the determination of the activated partial thromboplastin time (APTT) were compared with respect to their factor VIII:C and factor IX sensitivity for measurements of canine plasma. For this purpose, plasma with different levels of factor VIII:C or factor XI activity (100, 80, 70, 60, 50, 40, 30, 20, 15, and 10% [factor VIII:C: additionally 5%]) was prepared by mixing pool plasma with plasma of dogs with haemophilia A or B. Double measurements of three different sample mixtures were carried out for each activity level. The sensitivity of the reagents was measured first based on the ratios of the coagulation time to the 100% values. In addition, the single factor activity (F VIII:C/IX(X0.975)), whose accompanying APTT corresponded to the upper limit of the reference range (97.5%-quantile, n = 50) of the respective reagent, was determined graphically. The APTT reflected a decrease of factor IX activity generally more sensitive than a reduction of factor IX activity of an identical degree. Based on ratios distant differences respecting factor VIII:C and factor IX sensitivity were found between different reagents using two way analysis of variance (p < 0.05). Significant differences between various reagents were also found with respect to the F VIII:C(X0.975) and the F IX(X0.975). These corresponded to values between 27 and 50% or 32 and 64%, respectively, dependent on the reagent. As a result, the more sensitive reagents fulfilled the demands on the sensitivity of APTT in humans. Based on the latter criterion the highest sensitivity for both factors was found for the same reagent (Pathromtin) consisting of kaolin as a contact activator and human placental phospholipid. Respecting all proofed reagents, however, no relation was found between contact activator and single factor sensitivity.

