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Effect of analytic uncertainty of conventional and point-of-care assays of activated partial thromboplastin time on clinical decisions in heparin therapy.

The authors assessed the capability of assays of activated partial thromboplastin time (aPTT) for supporting clinical decision algorithms for heparin therapies of varying complexity. Blood samples were collected prospectively in three explicit management strategies from 100 sequential patients for whom heparin dosage was adjusted for therapeutic monitoring, femoral venous sheath removal after cardiac catheterization, or heparinization after thrombolytic therapy. In two- and three-way decision algorithms, conventional and point-of-care aPTT assays agreed with heparin assays in approximately two thirds of cases, and the two aPTT assays agreed in 80% or more of all cases. In six-way decision algorithms, the two aPTT assays agreed in only about half of all cases. The authors conclude that the reliability of point-of-care aPTT assays is similar to that of conventional assays. Both techniques can support two- and three-way decision algorithms but not some more complex patient classifications.

Algorithms↗

Prothrombin time and activated partial thromboplastin time can be performed on the first tube.

The current practice standard for coagulation studies on venous blood specifies the use of the second (or subsequent) tube for testing. Unless other tests are ordered, the first tube is discarded. This practice, derived from early experience using nondisposable materials, gained support from a report based on modern phlebotomy methods but inexperienced phlebotomists. For our study, paired blood specimens were collected by experienced phlebotomists from 175 outpatients whose physicians had ordered coagulation tests. Prothrombin time (PT) and activated partial thromboplastin time (APTT) were performed on the first and second tubes. Similar values were obtained for PT and APTT. (PT: n = 174; mean = 15.36 seconds; mean difference = 0.10 seconds; y = 1.02x - 0.28; t = 0.24; P>.81; r = 0.995; APTT: n = 160; mean = 38.25 seconds; mean difference = 0.48 seconds; y = 1.03x - 0.71; t = 0.25; P=.80; r = 0.993.) For coagulation tests on specimens collected by experienced phlebotomists, the first tube yields accurate results and need not be discarded.

Blood Coagulation Tests↗

Comparison of anti-factor Xa heparin activity and activated partial thromboplastin time in 2,773 plasma samples from unfractionated heparin-treated patients.

This study was conducted to evaluate two methods of determining the "therapeutic range" of the activated partial thromboplastin time (APTT) during unfractionated heparin treatment. Multiple fresh plasma samples from 694 patients treated with unfractionated heparin were tested simultaneously for APTT and anti-factor Xa heparin activity. The data were analyzed by linear regression and by a minimization-of-error technique to determine the more accurate APTT therapeutic range. The best-fit linear regression equation was obtained using logarithmically transformed APTT values. Using this equation, the heparin activity predicted by an APTT value had a 95% limit of agreement of +/- 0.39 U/mL compared with the actual heparin activity. The total clinical error rate of the APTT therapeutic range selected using the best-fit regression equation was 10%. An improvement to 8% was achieved using a method that chose the therapeutic range based on minimizing the APTT errors. Even 8% may be too high an error rate for best clinical results.

Evaluation Studies as Topic↗

Comparing different lots of activated partial thromboplastin time reagent: analysis of two methods.

This study was conducted to compare the performance of 2 different lots of activated partial thromboplastin time (APTT) reagent from the same manufacturer. Two hundred plasma samples were analyzed for APTT on the same instrument using 2 separate APTT reagent lots. Results were compared with those obtained from duplicate APTT determinations with 1 reagent lot on an additional 267 plasma samples. Regression analysis of paired APTT data was suboptimal because the disagreement of the results increased as the APTT values became higher. A better approach was the use of the Bland and Altman assessment of agreement method on logarithmically transformed data, which showed that the new reagent lot produced values that were 88.9% of the values produced by the old lot. This importantly reduced the APTT therapeutic range for unfractionated heparin (UH). In comparison, duplicate APTT values were 97.6% of a first APTT value. However, 95% limits of agreement for duplicate APTT determinations were +/-10% over the range of values considered therapeutic for UH. The variability of APTT results within and between reagent lots is one reason that APTT may not be optimum for monitoring UH therapy.

Anticoagulants↗

The activated plasma recalcification time. A Measurement encompassing the activated partial thromboplastin time and platelet function, with improved detection of bleeders.

A study of 300 patients is presented to demonstrate the effectiveness of the activated plasma recalcification time (APRT) as a measure of the intrinsic pathway and platelet function in the detection of bleeders. The activated partial thromboplastin time (APTT) and platelet count were determined in all patients having both normal and abnormal APRT's and the results correlated with overt bleeding tendencies. Abnormality of the APTT is closely paralleled by abnormality of the APRT, independent of the platelet count. When the APTT is normal, an abnormal APRT will characterize clinical bleeders with greater frequency than will the platelet count. The value of the APRT is greatest when done in concert with other standard coagulation measurements, as it both reinforces and expands their diagnostic capability.

Adolescent↗

Comparison of several activated partial thromboplastin time methods.

Activated partial thromboplastin times (APTT's) performed with a semi-automated electrical-conductivity type of clot timer on plasmas from patients with hepatic disease and intravascular coagulation, and on warfarin or heparin therapy, were significantly lower than when done on the same plasmas with either a manual optical method or an automated optical-endpoint instrument. Results of APTT's done on normal plasmas by the three methods were not significantly different. Substitution of different activator-phospholipid reagents resulted in some variability in results, but these differences were less than those between the different done with both the electrical clot timer and the automated optical instrument on prepared plasmas containing 5.0 or 1.0% of factor II, V, VIII, IX, OR X revealed shorter times with the electrical clot timer only in the case of factor II- and factor V-deficient plasmas. APTT's done on normal plasmas to which 0.1 or 0.3 units per ml. of heparin had been added vitro also were shorter with the electrical clot itmer than the automatic optical instrument. Prothrombin times done on normal and abnormal control plasmas and on a series of plasmas from patients on warfarin therapy showed no significant difference between the two methods.

Autoanalysis↗

Variable response of activated partial thromboplastin time to heparin therapy during hemodialysis.

The consistency of the anticoagulant effect of intravenously administered heparin was studied. The activated partial thromboplastin time (APTT) was measured for six patients hourly during three consecutive hemodialysis sessions each. Cephaloplastin was the plasma-activating agent. The time required to form a clot was measured by a light-sensitive electronic timer and confirmed within +/- 5% by the tilt tube method. Results are reported in second relative to units of heparin given to patients per kilogram body weight. The range of APTT's measured 55 minutes after each heparin dose greatly exceeded the range of technical variability of the assay method. The probably mechanisms and consequences for this variability after a constant heparin dose are discussed. The anticoagulation effect of heparin during hemodialysis in an otherwise stable clinical situation is not constant. The risks of having too much or too little anticoagulation are not eliminated by having determined a therapeutic heparin dose during one dialysis rung.

Blood Coagulation Tests↗

The effect of increased contact activation time on the activated partial thromboplastin time.

With the kaolin-cephalin activated partial thromboplastin time technic, the plasmas of persons who have Fletcher factor deficiency have shown considerable shortening of clotting times when contact activation has been lengthened from 3 (PTT-3) to 10 minutes (PTT-10). The authors demonstrate that in plasma of most normal individuals, and in coagulopathies of other sorts, only slight shortening usually occurs. Abnormal shortening occurs in plasmas of a few otherwise normal people, the "slow activators," and patients receiving coumarin drugs, who have greatly prolonged prothrombin times. Longer activation may produce greatly prolonged PTT's in plasmas containing heparin in relatively high concentrations. The authors discuss the significance of these findings.

Blood Coagulation Disorders↗

Heparin and the activated partial thromboplastin time--a difference between the in-vitro and in-vivo effects and implications for the therapeutic range.

The in-vivo sensitivity to heparin of activated partial thromboplastin time (aPTT) reagents is different from the in-vitro sensitivity. Equivalent therapeutic ranges for reagents of very different sensitivities may be determined from the regression line of aPTT against whole blood clotting time. The commonly used ratio of 1.5 to 2.5 times the normal control value is appropriate for some reagents, but for the other reagents a higher or lower range is appropriate. A suitable aPTT reagent for heparin control should be sensitive both to a threshold effect of heparin and to increasing dosage. Some reagents discriminate poorly at the threshold level.

Heparin↗

Fletcher factor deficiency, source of variations of the activated partial thromboplastin time test.

The activated partial thromboplastin time test measures the integrity of the intrinsic clotting system. The sensitivity of this test to Fletcher factor deficiency is dependent upon the particular protocol and reagents utilized in the test system. The case report presented here demonstrates conflicting laboratory results obtained for a patient who had Fletcher factor deficiency, the results depending on the laboratory choice of test reagents. This serves to emphasize the importance of physician awareness of the procedures utilized in laboratory testing.

Aged↗

Partial thromboplastin time in the presence of heparin: a rapid polybrene neutralization method.

The detection of heparin in plasma and its neutralization to permit assessment of the prolonged partial thromboplastin time is a constant need in the clinical laboratory. A rapid, simple method which utilizes Polybrene after the initial contact activation step is described and compared to the standard neutralization procedures. The present method has the advantage of not requiring Polybrene titration and of requiring only small volumes of plasma making it particularly valuable in pediatric patients.

Blood Coagulation Tests↗

Heparin monitoring by activated partial thromboplastin time. Comparison of ex vivo measurement and in vitro standardization.

Twenty patients were studied prospectively during heparin therapy. Three activated partial thromboplastin time (APTT) reagents were used to compare APTT values with plasma heparin levels during induction of heparin and transition of heparin to coumadin. A heparin in vitro dose APTT response curve and a heparin ex vivo curve were established. The in vitro sensitivity curves using different reagents were varied at therapeutic heparin levels. In contrast, the APTT reagents did not differ ex vivo. The in vitro curves demonstrated poor performance. Sixty percent of the patients did not adequately compare by APTT estimation of plasma heparin levels. An APTT ratio (1.5 to 2.5) using the patient's baseline APTT as the denominator demonstrated better representation of heparin levels. The in vitro APTT curves are inappropriate for heparin monitoring.

Adult↗

A comprehensive evaluation of the performance of duplicate prothrombin time and activated partial thromboplastin time assays.

An evaluation of the performance of duplicate prothrombin time (PT) and activated partial thromboplastin time (aPTT) assays was undertaken to develop analytical duplicate performance criteria in order to quantitate the risks associated with singlet versus duplicate procedures. Data were retrospectively collected from two hospital laboratories using two different coagulation systems. Included in the study were 6,391 patient samples; 3,047 PT, 3,334 aPTT, for a total of 12,782 data points. If a difference between duplicates of 5% or less is deemed analytically (or clinically) insignificant for PT, then fewer than 1% of the samples analyzed by either laboratory would require duplicates. If a difference between duplicates of 15% or less is deemed analytically (or clinically) insignificant for aPTT, then fewer than 2% of samples would exceed this limit for laboratory A, but 6.0% of samples from laboratory B exceeded this limit.

Blood Coagulation Tests↗

The variations between heparin sensitivity of different lots of activated partial thromboplastin time reagent produced by the same manufacturer.

The variations between different lots of activated partial thromboplastin time (APTT) produced by three major North American suppliers were evaluated over the past eight years. The authors found significant variations between the heparin sensitivity of the APTT reagents produced under the same name by the same supplier. The variations were so much that, using the recommended APTT ratio or prolongation of APTT for monitoring heparin therapy, one would have achieved significantly different intensity of heparinization from year to year.

Blood Coagulation Tests↗

Effect of clot-detection methods and reagents on activated partial thromboplastin time (APTT). Implications in heparin monitoring by APTT.

Two automatic coagulometers, ACL 810 (Instrumentation Laboratory), a laser-nephelometric centrifugal analyzer, and KoaguLab 40 A (Ortho Diagnostics), an optical automatic coagulometer, were compared with the manual tilt-tube method for the performance of activated partial thromboplastin time (APTT). Seven commercial APTT reagents were used for duplicate determinations in 30 normal controls, 26 patients with liver disease, and 33 patients on full-dose heparin treatment. Clotting times were longer with the manual method than with ACL 810 and, to a lesser extent, with KoaguLab 40 A. Average imprecision of duplicate determinations (coefficient of variation [CV]) was less with ACL 810 (less than 1.5%) than with KoaguLab 40 A (2.9%) and the manual method (2.4%). Differences in slope of the regression curves of clotting times obtained with the coagulometers over the tilt-tube method were observed with all the reagents tested (P less than 0.01). Transformation of clotting times of controls, patients with liver disease, and patients on heparin therapy to APTT ratios did not eliminate the bias resulting from the different reagents (P less than 0.001) and clot-detection methods (P less than 0.001); in controls, significant (P less than 0.001) reagent-method interaction was also observed. The in vitro heparin sensitivity differed with the APTT reagents evaluated and was influenced by the clot-detection method used. Transformation of APTT ratios of anticoagulated patients to apparent plasma heparin levels--as derived from in vitro dose-response curves--effectively eliminated the bias resulting from the different clot-detection methods but had no effect on the bias resulting from the different APTT reagents. In vitro heparin activity curves thus have little, if any, relevance for the ex vivo monitoring of heparin treatment.

Blood Coagulation Tests↗

An analysis of duplicate testing of prothrombin time and activated partial thromboplastin time assays.

An evaluation of duplicate prothrombin time (PT) and activated partial thromboplastin time (aPTT) assays determined by the MCA 110 coagulation analyzer was undertaken to develop analytical duplicate performance criteria to quantitate the risks associated with single versus duplicate procedures. Included in the study were 1,277 patient samples. On the basis of the currently recommended therapeutic range for prothrombin ratios, a variation of approximately 10% or more between duplicates was considered to be unacceptable. For aPTT assays, the recommended therapeutic range for heparin therapy was usually 1.5 to 2.5 times the baseline value, and variations of up to 25% might be considered acceptable. With these relatively lenient criteria, approximately 2% of PT and 1.3% of aPTT assays had differences between duplicate values that were unacceptable. From this data the authors concluded that the frequency of errors produced by single estimations was too great for satisfactory clinical practice.

Humans↗

Prognostic implication of activated partial thromboplastin time after reteplase or half-dose reteplase plus abciximab: results from the GUSTO-V trial.

AIMS: To evaluate the relationship between activated partial thromboplastin time (aPTT) and clinical outcomes in the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO-V) trial comparing standard-dose reteplase to half-dose reteplase and abciximab. METHODS AND RESULTS: We analysed data on 11,420 patients receiving unfractionated heparin. Peak aPTT levels recorded during the hospitalization were correlated with clinical outcomes. Multivariable logistic regression models examined the relationship between peak aPTT levels and (i) moderate-to-severe bleeding, (ii) intracerebral haemorrhage, (iii) reinfarction, and (iv) 30-day mortality. Non-linear relationships were explored in the models using cubic spline functions. Higher rates of significant complications were seen in both groups when aPTT levels were <50 s or when levels were >70 s. In the combination therapy group, the relationship between aPTT levels and bleeding appeared accentuated. Reinfarction rates increased gradually as aPTT levels were >70 s in both groups, but the relationships were not statistically significant. Peak aPTT levels <50 s were associated with increased 30-day mortality even after multivariable adjustment. CONCLUSION: Peak aPTT levels <50 s and >70 s are associated with worse clinical outcomes in the modern era of fibrinolytic therapy; these relationships are different in patients receiving standard reteplase vs. combination therapy.

Abciximab↗

A coupled amidolytic assay for thromboplastin (tissue factor) using a fluogenic substrate: its application to monkey leukocyte tissue factor.

A sensitive and quantitative amidolytic assay for thromboplastin (tissue factor) coupled to thrombin formation was established. A fluogenic peptide substrate, Boc-Val-Pro-Arg-MCA, was found to be suitable for the coupled amidolytic assay. The amidolytic assay was applied to measure TF activity of endotoxin-stimulated mononuclear leukocytes and monocytes. The amidolytic assay showed good correlation of 0.97 with the currently used clotting assay upon measuring TF activity of the cellular samples.

Animals↗