Multicentric evaluation of a new PT reagent based on recombinant human tissue factor and synthetic phospholipids

A new PT reagent based on recombinant human tissue factor and synthetic phospholipids (phosphatidyl choline and phosphatidyl serine) with defined fatty acid side chains was calibrated against BCT/253 and CRM 149R. A small but consistent bias in the International Sensitivity Index (ISI) value was obtained using either the human or rabbit brain reference material. ISI values were around 1.0 or slightly lower depending on the respective instrument. Mixing studies with factor deficient plasmas showed a high factor sensitivity especially for factor VII as compared to commercial rabbit brain or human placenta thromboplastin. In an international field trial the reagent was tested using fully or semi automated Electra® coagulometers in 4 different laboratories. Results with normal samples were in excellent agreement among the different laboratories. Mean values were 10.9, 10.9, 11.0, 11.2 s with a range of 9.5 to 12.5 s. Results of males and females were not different. In patients with liver disease very similar PT activities were found as compared to sensitive rabbit brain or human placental thromboplastins. In normals and patients with oral anticoagulation INR values correlated very well against BCT (r = 0.98, regression line y = -0.07 + 0.9 x). The distribution of samples was linear over the whole range. In the comparison against sensitive rabbit brain thromboplastin or human placental thromboplastin similar correlations were found. In a few cases higher INR values were observed for the recombinant reagent especially in patients with intensive treatment. Factor assays in those patients showed at least the strong reduction of one vitamin K-dependent coagulation factor. Over all the linearity was better against the rabbit brain reagent than against the human placental reagent which is slightly less factor VII sensitive as shown in mixing studies with normal and factor VII deficient plasma. Precision studies in the 4 laboratories showed excellent reproducibility of lyophilised controls or local patient plasma pools for all reagents with a better performance of the recombinant reagent. C.V. values from day to day ranged from 1.3% to 5% for normal and abnormal controls. These results show that the recombinant PT reagent, especially in conjunction with a precise automated instrument, may improve the results of PT testing and thus may lead to better patient care.