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PubMed2006

A national field study of quality assessment of CoaguChek point-of-care testing prothrombin time monitors.

A system for quality assessment (QA) of the CoaguChek (Roche Diagnostics, Mannheim, Germany) point-of-care testing prothrombin time monitor has been developed by the European Concerted Action on Anticoagulation. Hitherto there has not been an adequate rapid method for CoaguChek QA. Sets of 5 certified international normalized ratio (INR) plasma samples were tested on 539 CoaguChek monitors by experienced staff at 9 Netherlands Thrombosis Centers and results compared with certified INR. A 15% or more deviation has been classified as significant deviation. Overall mean and certified INR values were similar, but 20.3% of participants showed a 15% or more deviation from the certified INR on at least 1 of the 5 QA plasma samples. Statistically significant differences in results with different lots of CoaguChek test strips were found. There is need for large scale QA of CoaguChek monitors. The importance of the 5 CoaguChek certified INR QA plasma samples being tested on a single occasion is demonstrated.

International Normalized Ratio↗

PubMed2004

Effect of direct thrombin inhibitors, bivalirudin, lepirudin, and argatroban, on prothrombin time and INR values.

Direct thrombin inhibitors (DTIs) represent a new class of promising anticoagulation agents. The DTIs frequently are used to provide initial anticoagulation, with long-term therapy requiring eventual transition to coumarins. Unfortunately, DTIs not only prolong the activated partial thromboplastin time but also can affect international normalized ratio (INR) values. We approximated the DTI effect on INRs by each drug to pooled plasma at concentrations between 0.1 and 1.2 microg/mL. We then concurrently tested these samples using 14 prothrombin time (PT) reagents. By using repeated measures analysis of variance, we found significant differences (P < .05) between the median INRs for lepirudin and argatroban for all PT reagents, between lepirudin and bivalirudin for all reagents except PT-Fibrinogen HS Plus (P = .07), and between bivalirudin and argatroban for all reagents except Thromborel S (P = .05). The DTI effect on INRs was dependent on drug, drug concentration, and reagent. Argatroban had the most effect on INRs, while lepirudin had the least effect. Reagents with a lower international sensitivity index were less affected by DTI; ThromboMax HS was the least sensitive PT reagent to any DTI.

Antithrombins↗

PubMed2001

Point-of-care prothrombin time measurement for professional and patient self-testing use. A multicenter clinical experience. Oral Anticoagulation Monitoring Study Group.

We enrolled 386 subjects in a multicenter study of a point-of-care (POC) prothrombin time (PT) testing device. POC tests were performed by health care professionals using venous and finger-stick specimens and by patients using finger-stick specimens. Venous blood also was analyzed in the local hospital laboratory and a national reference laboratory. Accurate POC results were obtained by professionals using both types of specimens. Patients' results were equivalent to those of professionals. The identification of the patient's therapeutic status based on the International Normalized Ratio (INR) was equivalent for POC and local hospital laboratory PT results; 75% of local laboratory results and 77% of POC results were within 0.4 INR of reference laboratory results, while 93% of either system (POC or local laboratory) were within 0.7 INR. Patients overwhelmingly reported satisfaction with the self-test, including the finger stick and device operation. The INR from the POC device is clinically equivalent to the laboratory INR for assessment of anticoagulation status and management decisions in professional and self-testing environments. Patients can learn to perform accurate PT testing, and POC PT testing is feasible in patients' homes.

Adolescent↗

PubMed2002

Point-of-care testing for prothrombin time, but not activated partial thromboplastin time, correlates with laboratory methods in patients receiving aprotinin or epsilon-aminocaproic acid while undergoing cardiac surgery.

Point-of-care testing (POCT) of coagulation parameters can help optimize transfusion practice in cardiac surgery. Antifibrinolytic agents may interfere with the laboratory and/or POCT coagulation assays. This randomized controlled study compared coagulation parameters obtained from a whole blood POCT coagulation device with a typical laboratory instrument in cardiac surgery patients receiving aprotinin, epsilon-aminocaproic acid, or normal saline before undergoing cardiopulmonary bypass. Aliquots of arterial blood samples from 42 patients were collected perioperatively, and their prothrombin times (PTs) and activated partial thromboplastin times (aPTTs) were measured by POCT and laboratory instrumentation. Linear regression and error analyses were used for the method comparison. For PT, the POCT device compared favorably with the laboratory method. For aPTT, the POCT device did not compare well with the laboratory method. Treatment with antifibrinolytic agents does not interfere with determination of PT.

Aminocaproic Acid↗

PubMed1999

The effect of warfarin therapy on the charge properties of urinary prothrombin fragment 1 and crystallization of calcium oxalate in undiluted human urine.

Urinary prothrombin fragment 1 (UPTF1) is the principal protein in calcium oxalate (CaOx) crystals precipitated from human urine and is a potent inhibitor of CaOx crystallization, a property that should depend, at least in part, upon the extent of gamma-carboxylation of the 10 glutamic residues in its N-terminal region. Warfarin therapy limits full gamma-carboxylation of vitamin K-dependent proteins, including UPTF1. The aims of this study were to determine the effect of warfarin therapy on UPTF1, its occlusion into CaOx urinary crystals, and its influence on the crystallization of CaOx in undiluted human urine. In the first part of the study, urines were collected from six men prior to cardiac surgery and after stabilization on long-term warfarin treatment. Proteins in the urines and in the matrix of CaOx crystals precipitated from them were analyzed by two-dimensional SDS-PAGE and Western blotting. In urine, at least two charge variants of UPTF1 with low isoelectric point (pI) values were detected before and during warfarin therapy, but additional higher pI forms of the protein were also seen during anticoagulation. Nonetheless, the majority of UPTF1 was present in the more fully gamma-carboxylated state. CaOx crystals precipitated from the same urine samples contained only low pI forms of UPTF1. The effect of warfarin treatment on CaOx crystallization in urine was tested by collecting two consecutive 24-h urine samples from 16 men prior to cardiac surgery and during subsequent warfarin treatment. CaOx crystallization was induced in each sample by the addition of sodium oxalate. The size and volume of the particles deposited were determined using a Coulter counter, and the crystals were examined by scanning electron microscopy (SEM). There were no significant differences between the urinary metastable limits before or during warfarin treatment or in the total volume of crystals precipitated. A slight increase in the mean diameter of the crystalline particles precipitated from the urines during anticoagulant therapy was not significant. SEM showed little evidence of changes in overall particle size, although individual crystals of CaOx tended to be larger during warfarin treatment. It was concluded from these studies that the binding of UPTF1 to CaOx crystal surfaces is related to the degree of gamma-carboxylation of its Gla domain, which would also influence the protein's inhibitory effects on CaOx crystallization. However, during warfarin therapy the majority of UPTF1 exists in a highly charged state, indicating that it is completely, or almost completely, gamma-carboxylated, which would explain the lack of any difference between CaOx crystallization parameters in the urine of subjects before and during warfarin administration. We conclude that physiologically significant reductions in the inhibitory potency of UPTF1 would be likely to occur only as a result of proscription of gamma-carboxylation more extensive than that induced by warfarin.

Adult↗

PubMed2004

European concerted action on anticoagulation. Quality assessment of the CoaguChek Mini and TAS PT-NC point-of-care whole-blood prothrombin time monitors.

BACKGROUND: International Normalized Ratios (INRs) for prothrombin time obtained with the CoaguChek Mini and TAS (RapidPointCoag) PT-NC systems are markedly different and also differ from the "true" INR. There is therefore a need for local quality assessment (QA) of the two systems. METHODS: A set of 60 lyophilized artificially depleted and 60 lyophilized coumarin plasmas were tested at 10 centers on both point-of-care testing monitors. Subsets of three and five plasmas were selected as QA plasmas and compared with the remaining 55 to assess the relative ability of the systems to characterize performance at the individual centers. The incidence of aberrant results (outliers; >15% deviation from the true INR) was also recorded. The expected incidence with the QA plasmas was calculated and compared. RESULTS: On both systems, INR with the common sets of 55 lyophilized plasmas varied considerably between centers. With the TAS PT-NC, subsets of five and three European Concerted Action on Anticoagulation (ECAA) artificially depleted plasmas gave good correlation with the 55 plasmas, but the coumarin plasmas performed less well. With the CoaguChek Mini, correlation was good with sets of five artificially depleted QA plasmas and reasonable with three but was less satisfactory with the coumarin plasmas. Outliers were detected with both types of plasmas on both test systems but with variable success. CONCLUSIONS: With the TAS PT-NC, three ECAA artificially depleted lyophilized plasmas provided reliable QA, but five lyophilized coumarin plasmas were required. With the CoaguChek Mini, five artificially depleted plasmas gave reliable QA but coumarin plasmas gave poorer results. ECAA QA plasmas provide a local system for checking INRs obtained with monitors of both types.

Anticoagulants↗

PubMed2005

Preanalytical variables and off-site blood collection: influences on the results of the prothrombin time/international normalized ratio test and implications for monitoring of oral anticoagulant therapy.

BACKGROUND: The quality of oral anticoagulant therapy management with coumarin derivatives requires reliable results for the prothrombin time/International Normalized Ratio (PT/INR). We assessed the effect on PT/INR of preanalytical variables, including ones related to off-site blood collection and transportation to a laboratory. METHODS: Four laboratories with different combinations of blood collection systems, thromboplastin reagents, and coagulation meters participated. The simulated preanalytical variables included time between blood collection and PT/INR determinations on samples stored at room temperature, at 4-6 degrees C, and at 37 degrees C; mechanical agitation at room temperature, at 4-6 degrees C, and at 37 degrees C; time between centrifugation and PT/INR determination; and times and temperatures of centrifugation. For variables that affected results, the effect of the variable was classified as moderate when <25% of samples showed a change >10% or as large if >25% of samples showed such a change. RESULTS: During the first 6 h after blood collection, INR changed by >10% in <25% of samples (moderate effect) when blood samples were stored at room temperature, 4-6 degrees C, or 37 degrees C with or without mechanical agitation and independent of the time of centrifugation after blood collection. With one combination of materials and preanalytical conditions, a 24-h delay at room temperature or 4-6 degrees C had a large effect, i.e., changes >10% in >25% of samples. In all laboratories, a 24-h delay at 37 degrees C or with mechanical agitation had a large effect. We observed no clinically or statistically relevant INR differences among studied centrifugation conditions (centrifugation temperature, 20 degrees C or no temperature control; centrifugation time, 5 or 10 min). CONCLUSIONS: We recommend a maximum of 6 h between blood collection and PT/INR determination. The impact of a 24-h delay should be investigated for each combination of materials and conditions.

Administration, Oral↗

PubMed2005

Poor agreement among prothrombin time international normalized ratio methods: comparison of seven commercial reagents.

BACKGROUND: Prothrombin time (PT) has long been the most popular test for monitoring oral anticoagulation therapy. The International Normalized Ratio (INR) was introduced to overcome the problem of marked variation in PT results among laboratories and the various recommendations for patient care. According to this principle, all reagents should be calibrated to give identical results and the same patient care globally. This is necessary for monitoring of single patients and for application of the results of anticoagulation trials and guidelines to clinical practice. METHODS: We took blood samples from 150 patients for whom oral anticoagulation had been prescribed. Plasmas were separated and PTs determined by use of seven commercial reagents and four calibrator sets. The differences in results were assessed by plotting, for each possible pair of methods, the differences in INR values for each sample against the mean INR value (Bland-Altman difference plots). RESULTS: Mean results differed significantly (P <0.001) for 17 of 21 possible paired comparisons of methods. Only two pairs of methods produced very similar results when assessed for problems of substantial differences in INR values; a significant, systematic increase in the difference with INR; and a significant systematic increase in the variation in difference with increasing INR values. CONCLUSIONS: The agreement among several (and perhaps most) commercial INR methods is poor. The failure of current calibration strategies may severely compromise both the monitoring of individual patients and the application of oral anticoagulation guidelines and trial results to clinical practice.

Administration, Oral↗

PubMed2006

Quality assessment of CoaguChek point-of-care prothrombin time monitors: comparison of the European community-approved procedure and conventional external quality assessment.

BACKGROUND: There is a need for dependable quality assessment (QA) of the widely used CoaguChek point-of-care testing prothrombin time monitor. By use of the prescribed set of 5 CoaguChek certified international normalized ratio (INR) QA plasmas, we compared the reliability of the immediate QA of individual monitors described in the European Community-recommended Technology Implementation Plan with conventional external QA analysis. METHODS: Experienced staff tested CoaguChek point-of-care monitors in routine use for controlling oral anticoagulant dosage at 9 Netherlands Thrombosis Service Centres. Testing was performed with both the certified CoaguChek INR for a set of 5 QA individual plasmas from the Eur Con Action on Anticoag (ECAA) and conventional external QA analysis. RESULTS: Patients brought 523 CoaguChek monitors to our service centers for assessment. The proportion with unsatisfactory performance indicated by a 15% deviation from the ECAA set was compared with 15% deviation from overall median INR of all CoaguChek monitors in the survey, as in conventional QA analysis. The results were similar (20.3% and 18.5%, respectively). Interlot differences of CoaguChek test strips were detected, but the incidence of unsatisfactory performance was similar with both analyses, from 6.5% to 37.5% with the certified INR method and from 5.9% to 33.3% with the overall median analysis. CONCLUSIONS: The results validate the use of the European Action on Anticoagulation rapid single-instrument QA-specific procedure for CoaguChek users compared with the nonspecific conventional QA analysis that relies on deviation from the overall median INR.

Administration, Oral↗

PubMed1997

Using the international normalized ratio to standardize prothrombin time.

The international normalized ratio, or INR, was introduced in 1983 by the World Health Organization, or WHO, Committee on Biological Standards to more accurately assess patients receiving anticoagulation therapy. The INR mandates the universal standardization of prothrombin time. This article describes the method used to calculate INR, as well as its clinical relevance to the practice of dentistry.

Anticoagulants↗

PubMedDate not supplied

Changes in prothrombin and activated partial thromboplastin time during replacement therapy with human recombinant growth hormone in growth hormone deficient adults.

BACKGROUND: In rodents, Growth Hormone (GH) has been shown to stimulate coagulation parameters, including Prothrombin Time (PT), activated Partial Thromboplastin Time (aPTT) and vitamin K dependent coagulation factors. However, there are no reports on the influence of GH replacement therapy on global coagulation tests in Growth Hormone Deficiency (GHD). OBJECTIVE: The aim of this study was to investigate the effects of GH administration on basic coagulation parameters: PT, aPTT and fibrinogen concentrations in adult GHD patients before and during one year of GH replacement. DESIGN: Twenty-one adult patients with severe GHD (mean age +/- SE: 38.6 +/- 2.8 years) were included in this hospital based, prospective, interventional study. All patients were treated with rhGH for 12 months (GH dose: 0.4 mg/day for male and 0.6 mg/day for female patients). IGF-1 concentrations were determined using RIA-INEP kits. Basic coagulation tests, i.e. aPTT and fibrinogen concentrations, were measured before and after 3, 6 and 12 months of treatment with rhGH. Control values were obtained from fourteen "healthy" subjects matched by age, sex and body mass index (BMI). RESULTS: At baseline, we observed no significant differences in PT, aPTT and fibrinogen values between GHD and healthy subjects. IGF-1 concentrations increased significantly within 3 months of GH therapy (8.2 +/- 1.5 vs. 24.2 +/- 2.9 nmol/l, p <0.05) and remained stable thereafter. A significant increase in PT values, which was more pronounced in female subjects, was noted after 6 and 12 months of treatment with GH. aPTT values increased significantly after 12 months of treatment only in male patients (28.8 +/- 4.6 vs. 39.7 +/- 2.1 s.; p <0.05). No significant changes in fibrinogen concentrations were found during the study. CONCLUSIONS: Twelve months of GH replacement therapy led to a significant increase in PT and aPTT values in adult GHD patients, while fibrinogen concentrations did not change. Changes in PT were more pronounced in female GHD patients, while an increase in aPTT values was observed only in male patients with GHD. The clinical significance of these changes needs further evaluation.

Adult↗

PubMed1989

Determination of haemoglobin and prothrombin complex in whole blood using optothermal spectrometry.

Optothermal spectrometry measures the thermal energy produced as a result of absorption by molecules at a given modulation frequency and wavelength. Depending on the modulation frequency used, analysis can be performed in very thin layers (50-150 micrometers). A major advantage of optothermal spectrometry is that it is not very sensitive to light scattering. Haemoglobin in whole blood was measured without any reagent at 16 Hz and 2 Hz frequencies. The precisions (within-series, within-day and between-day) were acceptable, and comparisons with reference methods were excellent. As opposed to the reference methods, optothermal spectrometric determinations were not affected by lipaemia. When measured continuously, the signal for haemoglobin increased due to the packing of erythrocytes towards the light source. By converting soluble fibrinogen into insoluble fibrin, the erythrocyte packing could either be inhibited or stopped, and this process could be monitored by assessing the change in the rate of signal increase for haemoglobin. This principle was utilized to analyse the prothrombin complex; the method was found to have acceptable precision and to be comparable to a routine method.

Blood Coagulation Factors↗

PubMed2002

Comparison of quick and owren prothrombin time with regard to the harmonisation of the International Normalised Ratio (INR) system.

Prothrombin time (PT) is tested mostly to monitor patients on oral anticoagulant treatment. The International Normalised Ratio (INR) was introduced to improve and harmonise PT results and therapeutic range globally for patient care and the scientific literature. We studied the Quick PT in 179 patients and the Owren PT in 137 patients on oral anticoagulant therapy using two different reagents for the two methods of measuring PT. We assessed the clinical significance of the INR results obtained by each method using the two reagents and compared the Quick and Owren methods. We conclude that with the Quick method individual INR results differed from each other too much clinically, while using the Owren method individual INR results were clinically acceptable. Our opinion is that we should develop the INR system using the Owren PT method rather than the Quick to improve patient care.

Administration, Oral↗

PubMed2002

Synergism between urinary prothrombin fragment 1 and urine: a comparison of inhibitory activities in stone-prone and stone-free population groups.

South African blacks rarely form kidney stones compared with whites. This study investigated whether purified urinary prothrombin fragment 1 (UPTF1) derived from blacks is a more potent inhibitor of calcium oxalate crystallisation than that from whites. UPTF1 was purified from the urine of both population groups and their inhibitory activities were compared in a cross-over design in which each protein was tested in ultrafiltered urine from both population groups. Coulter Multisizer, [14C]-oxalate deposition and scanning electron microscopy experiments were used to monitor crystallisation. The study has demonstrated for the first time that UPTF1 promotes nucleation and that inhibitory activity is synergistically dependent upon urine composition. The activity of the whites' UPTF1 was greater than that of the blacks in the whites' urine (e.g. particle size decrease: 31.7% vs. 25.2%), while the blacks' UPTF1 was superior to that of the whites in the blacks' urine (e.g. particle size decrease: 46.5% vs. 32.4%). In addition, when tested in their respective endogenous urines, the blacks' UPTF1 demonstrated superior inhibitory activity on an absolute scale (e.g. particle size decrease: 46.5% vs. 31.7%). Thus, the urine composition of black South Africans may influence their UPTF1 conformation, conferring greater efficacy for inhibition of calcium oxalate crystallisation.

Black People↗

PubMed2005

Reference materials (RMs) for analysis of the human factor II (prothrombin) gene G20210A mutation.

The Scientific Committee of Molecular Biology Techniques (C-MBT) in Clinical Chemistry of the IFCC has initiated a joint project in co-operation with the European Commission, Joint Research Centre, Institute of Reference Materials and Measurements to develop and produce plasmid-type reference materials (RMs) for the analysis of the human prothrombin gene G20210A mutation. Although DNA tests have a high impact on clinical decision-making and the number of tests performed in diagnostic laboratories is high, issues of quality and quality assurance exist, and currently only a few RMs for clinical genetic testing are available. A gene fragment chosen was produced that spans all primer annealing sites published to date. Both the wild-type and mutant alleles of this gene fragment were cloned into a pUC18 plasmid and two plasmid RMs were produced. In addition, a mixture of both plasmids was produced to mimic the heterozygous genotype. The present study describes the performance of these reference materials in a commutability study, in which they were tested by nine different methods in 13 expert laboratories. This series of plasmid RMs are, to the best of our knowledge, the first plasmid-type clinical genetic RMs introduced worldwide.

Base Sequence↗

PubMed2006

Factor V Leiden, prothrombin G20210A substitution and hormone therapy: indications for molecular screening.

Venous thromboembolism is a well-known complication of oral contraception and hormonal replacement therapy. Inherited thrombophilia is viewed as an important determinant in modulating the effects of estrogens on thrombotic risk. An increasing number of kits for thrombophilic mutations [factor V Leiden, G20210A prothrombin and methylenetetrahydrofolate reductase (MTHFR) C677T genes] are becoming commercially available, and screening for inherited thrombotic risk is among the most requested genetic tests in molecular diagnostic laboratories. However, the question of routine genetic screening for thrombophilia before prescribing hormones is still a matter of debate. The purpose of this article is to discuss the usefulness and practical applications of thrombotic genetic testing to identify which women should be tested to improve both the safety and efficacy of individualized estrogen therapy.

Animals↗

PubMed2006

A modified method of prothrombin time/International Normalised Ratio determination in capillary blood and monitoring oral anticoagulant therapy.

BACKGROUND: Oral anticoagulant therapy is monitored by a prothrombin time (PT) assay. The PT is standardised by the International Normalised Ratio (INR). The purpose of this study was to work out a modified method of PT/INR measurement in capillary blood for monitoring anticoagulation treatment. METHODS: Healthy donors, subjects with high or low haematocrit values, and oral anticoagulant-treated patients were included in the study. Plasma and capillary blood PT/INRs were determined by the standard Quick clotting assay, by the modified approach and with the CoaguChek S analyser. RESULTS: The performance characteristics of the developed method were accuracy, due to taking into account whole capillary blood haematocrit values, and precision, due to a decrease in the viscosity of the analysed samples. Implementation of the modified method showed that it is possible to use PT values of normal plasma for capillary blood INR calculation. The developed method allowed the determination of PT in capillary blood within the haematocrit value range from 0.15 up to 0.7. For capillary blood, the results of the modified method closely correlated with PT/INR values determined by the reference Quick method in venous plasma (r=0.99) and with the CoaguChek S analyser (r=0.97). CONCLUSIONS: The modified method of capillary blood PT/INR determination could be recommended for oral anticoagulant therapy monitoring.

Administration, Oral↗

PubMed2005

Barriers to patient self-testing of prothrombin time: national survey of anticoagulation practitioners.

Oral anticoagulation with warfarin requires routine monitoring of prothrombin time, expressed as the international normalized ratio (INR). Patient self-testing for INR is common in Europe but not in the United States. In order to determine the frequency of INR self-testing among patients whose anticoagulant therapy is managed in U.S. anticoagulation clinics, to describe the processes that support this self-testing, and to identify the barriers as experienced by anticoagulation clinic providers, a three-part survey was mailed to 538 anticoagulation specialists in the United States. The response rate was 43.7%. Policies and procedures of almost 60% of anticoagulation clinics prohibited INR self-testing for enrolled patients. In addition, less than 1% of patients being managed by U.S. anticoagulation clinics use self-testing to obtain INR results. Primary barriers were the cost of self-testing instruments (78.7% of respondents), cost of reagent cartridges (60.4%), and fear that self-testing might lead to unintended self-management (35.7%). Over 75% of respondents believed that some reimbursement for the cost of self-testing devices and supplies would increase the likelihood that anticoagulation clinics would recommend INR self-testing.

Ambulatory Care Facilities↗

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