Summary The prothrombin time (PT), obtained from a fresh normal plasma pool (FPP), is the basis both for the establishment of the 100% activity (normal plasma) and for the ratio calculation used in the International Normalized Ratio (INR) according to the recommendations of the ICSH/ICTH (6). Today the PT of lyophilized normal plasma pools are successfully used as reference for the assessment of samples in proficiency studies. However, a lack of comparability is to be recognized. Therefore the Committee of Hematology of the German Association of Diagnostics’ and Diagnostic Instruments’ Manufacturers (VDGH) decided to produce a candidate reference plasma (VDGH Reference Plasma) which was calibrated against fresh normal plasma pools in an international study. The basic calibration was performed by using the same certified BCR thromboplastin (BCT/099) by all participants. The endpoint was determined manually and by using the coagulometer Schnitger-Gross. In additional testings each participant used his own routine thromboplastins and methods. Calculating the ratio [PT VDGH Reference Plasma (sec)/PT fresh normal plasma pool (sec)] the VDGH Reference Plasma showed a deviation from the average fresh normal plasma pool of 1.05 both with the BCT/099 and with all thromboplastins. There were obtained some statistical differences between “plain” and “combined’’ (added factor V and fibrinogen) thromboplastins. No statistical difference was found between the different endpoint measurement methods (manual, mechanical, optical). In spite of these statistical deviations the VDGH Reference Plasma can be used for the standardization of the PT-normal (100%) value with different ratios for plain (1.06) and combined (1.02) thromboplastins. The manufacturers will use this VDGH Reference Plasma for the calibration of their commercially available calibration plasmas, which allows the user of such a material to calculate a calibrated 100% PT value.
Monitoring of direct thrombin inhibitors (DTIs) in patients with heparin-induced thrombocytopenia (HIT) is primarily performed using the activated partial thromboplastin time (aPTT). This assay is poorly standardized, reagent dependent, and not DTI specific. We compared aPTT, thrombin time (TT), and prothrombin time (PT) to drug levels obtained by the ecarin chromogenic assay (ECA). We analyzed 495 samples of patients with confirmed or suspected HIT on treatment with either argatroban (n = 37) or lepirudin (n = 80). Mean DTI levels ± standard deviation (SD) were 0.41 ± 0.36 µg/mL for argatroban and 0.20 ± 0.21 µg/mL for lepirudin. Results of aPTT were highly variable: 67 ± 22 seconds for argatroban and 55 ± 20 seconds for lepirudin. Significant correlations ( P < .01) were found between ECA-based DTI level and TT (argatroban, r = .820 and lepirudin, r = .830), PT (argatroban, r = -.544), and aPTT (lepirudin, r = .572). However, there was no correlation of aPTT with argatroban or PT with lepirudin concentration. Multiple regression analyses revealed that the TT predicted 54% of argatroban and 42% of lepirudin levels, but no significant impact was seen for PT or aPTT. The aPTT-guided monitoring of DTI therapy leads to a high percentage of patients with inaccurate plasma levels, hence resulting to either undertreatment or overtreatment. Knowledge of baseline values prior to DTI therapy and inclusion of clinical settings are essential for dosing DTIs when using aPTT. However, due to several limitations of aPTT, monitoring according to exact plasma concentrations as obtained by specific tests such as ECA may be more appropriate.
A multicenter study of a chromogenic substrate method for photometric determination of prothrombin time was conducted in order to evaluate its clinical application. Seven laboratories participated in the study using a total of 742 plasma samples from 417 patients on oral anticoagulant therapy, 261 healthy subjects and 64 patients with different diseases especially of the liver as well as 30 patients with hereditary deficiency of coagulation factors II, V, VII, X. The chromogenic PT method was compared to a standardized coagulometric PT assay which uses the same sensitive human placenta thromboplastin calibrated against international reference preparations. A high correlation of the prothrombin ratio values of the chromogenic and the coagulometric assay was obtained in 402 plasma samples (r = 0.940; y = 1.02x - 0.1). The study showed that the chromogenic PT reagent is sensitive to deficiency of the coagulation factors of the extrinsic pathway but not affected by heparin up to 1 IU/ml because of the heparin antagonist added. The precision (coefficient of variation) of the photometric method ranged between 0.6 and 3% (intraassay CV) and between 1.4 and 5.8 (interassay CV). The International Sensitivity Index (ISI) obtained for the used lot was 1.09. The therapeutical range in percentage activity for patients in a stable phase of an anticoagulant therapy was found to be from 15 to 27 percent of normal. The results of the clinical evaluation proved the good comparability of the new chromogenic PT test with coagulometric methods, its high factor sensitivity, good reproducibility and easy performance.
Article Evaluierung eines neuartigen Thromboplastins auf der Basis von rekombinantem humanem Tissue Factor und synthetischen Phospholipiden was published on January 1, 1993 in the journal Journal of Laboratory Medicine (volume 17, issue 11).
Introduction The T-TAS® (Total Thrombus formation Analysis System) is a device for assessing hemostasis in whole blood under physiological flow conditions. Resembling a modified flow chamber principle, it measures the change of flow during thrombus formation within micro capillaries in disposable microchips. The global assays measure different aspects in hemostasis. We determined the normal ranges of the three available chip types for a typical European healthy population and compared the variability between labs.
Abstract Background Light transmission aggregometry (LTA) is considered the gold standard for the evaluation of platelet function but is labor-intensive and involves numerous manual steps. Automation may contribute to standardization. Here, we evaluate the performance characteristics of a new automated instrument, Thrombomate XRA (TXRA), and compare it against a manual instrument (PAP-8). Materials and Methods Leftover blood samples from blood donors or patients were tested in parallel with identical reagents and in identical concentrations both manually using PAP-8 and automated on the TXRA. In addition to precision and method comparison, an additional evaluation was performed on the TXRA against “virtual” platelet-poor plasma (VPPP) based on artificial intelligence. The main focus was on comparing the maximum aggregation (MA%) values. Results Precision for MA% ranged from 1.4 to 4.6% on TXRA for all reagents. Normal ranges for 100 healthy blood donors on both instruments were in a similar range for all reagents, with a tendency to slightly higher values with TXRA. Most agonists resulted in normally distributed MA%. Comparing 47 patient samples on both devices showed a good correlation for both slope and MA% with some differences in individual samples with epinephrine and TRAP. Correlation between the TXRA measurement against PPP and “virtual” PPP demonstrated excellent correlation. Reaction signatures of both devices were very similar. Conclusion TXRA provides reproducible LTA results that correlate with an established manual method when tested against PPP or VPPP. Its ability to perform LTA only from platelet-rich plasma without requiring autologous PPP simplifies LTA. TXRA is an important step not only for further standardizing LTA but also for a more widespread use of this important method.
