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Decentralized testing for prothrombin time and activated partial thromboplastin time using a dry chemistry portable analyzer.

Previous work has established the precision and accuracy of a portable blood coagulation analysis system using paramagnetic particles contained in a dry reagent on a disposable test card. We examined the deployment of this technology in decentralized hospital settings and compared test results obtained in the surgical intensive care unit, coronary care unit, and outpatient cardiology clinic with those obtained in the central laboratory. Nursing personnel were instructed in the use of the system, and quality control testing was performed daily by the laboratory staff. In the intensive care units, patient subjects included those on whom tests of prothrombin time and activated partial thromboplastin time had been ordered. Immediate determinations were performed by the intensive care unit nursing staff on the same citrated, whole-blood samples that were subsequently sent to the central laboratory. In the outpatient cardiology clinic, fingerstick blood samples were obtained for prothrombin time determinations with the dry chemistry system. Paired prothrombin time samples obtained by venipuncture were run in the hospital laboratory. The study involved multiple users, multiple locations, two lots of activated partial thromboplastin time cards, and several different instruments, over an extended period. Correlation coefficients between the dry chemistry system and the hospital laboratory under these conditions were in an acceptable range in all sites studied. We concluded that, with appropriate training and quality assurance, the dry chemistry system provides an acceptable alternative to the hospital laboratory for prothrombin time and activated partial thromboplastin time determinations.

Chemistry Techniques, Analytical↗

[Control of the treatment with oral anticoagulants and prothrombin time].

There are many patients on oral anticoagulant therapy monitored with prothrombin time (Quick Test). Differences in sensitivity of the various types of tissue thromboplastins are the cause of the different results and common therapeutic ranges, calibrated reference thromboplastins with a defined International Sensibility Index (ISI) have become available. The ISI is used to estimate the International Normalized Rate (INR) which is used as a universal scale for the expression of the prothrombin time results in the control of oral anticoagulant therapy.

Administration, Oral↗

Effect of melagatran on prothrombin time assays depends on the sensitivity of the thromboplastin and the final dilution of the plasma sample.

Prothrombin time (PT) assays are clotting methods that measure the activity of vitamin K-dependent coagulation factors (F) II, VII, and X. There are three main types of PT assays in general usage, namely the Quick assay, Owren's assay and PT dry chemistry test cards. PT assays were initially developed to monitor dose-adjustments of vitamin K antagonists such as warfarin. The aim of the present study was to investigate whether commercially available PT assays are suitable for evaluating the anticoagulant activity of direct thrombin inhibitors. Melagatran, a reversible direct thrombin inhibitor, was added to human plasma at concentrations ranging from 0.1 to 2.0 micromol/l. Seventeen different commercially available PT kits were used, including thirteen Quick reagents, two Owren reagents and two PT test cards. The sensitivity of the different reagents, expressed as the concentration of melagatran that doubled the prothrombin time (IC50) varied widely, with Thromboplastin S and Thromboplastin HS being the most sensitive (IC50 = 0.9 micromol/l). The reagents with apparently the lowest sensitivity were the two Owren reagents Nycotest PT and SPA 50 with an IC50 of 2.2 and 2.9 micromol/L, respectively. This is most likely due to a higher dilution of melagatran in these assays compared to the dilution in the Quick assays. The results were also dependent on the International Sensitivity Index (ISI) of each reagent. The concentration of melagatran that produced an International Normalized Ratio (INR) of 2 was calculated from dose-response curves for each assay, and these results revealed that reagents with a high ISI value gave an INR of 2 at much lower concentrations of melagatran (0.5-0.7 micromol/L) than those with an ISI-values around one (0.9-1.2 micromol/L). It was found that INR depends not only on the plasma concentration of melagatran, but also on the sensitivity of the PT reagent and on the final dilution of the plasma sample in the prothrombin time assay. Thus, since the same melagatran concentration can be associated with widely varying PT/INR results depending on the specific assay used it is concluded that PT assays and INR can not be used to monitor melagatran activity.

Anticoagulants↗

Comparison of liquid and dried sodium citrate as the anticoagulant for Thrombotest and prothrombin time estimations.

Samples which are taken for the Quick one-stage prothrombin time estimation for the control of patients receiving oral anticoagulant treatment are by convention taken into liquid sodium citrate anticoagulant. Dried sodium citrate has been rejected on the grounds that excessive haemolysis causes activation of clotting factors and therefore falsely short clotting times. Tests were performed with both liquid and dried sodium citrate as an anticoagulant for prothrombin time and Thrombotest. No clinically significant difference between the values given by the two anticoagulants was observed.

Anticoagulants↗

Polycythaemia vera and Christmas disease with slightly prolonged prothrombin time.

A family is reported in which Christmas disease in one member and polycythaemia in another were associated with prolongations of the one-stage prothrombin time of the factor-VII deficiency type. Changes in the prothrombin time were present in the siblings of the case of Christmas disease and in a number of unrelated cases of polycythaemia vera. The use of "mixing procedures" did not reveal any significant mutual correction of the changes in prothrombin time.

Hemophilia A↗

Effects of haemolysis, lipaemia and bilirubinaemia on prothrombin time, activated partial thromboplastin time and thrombin time in plasma samples from healthy dogs.

Interferences caused by haemolysis, lipaemia and bilirubinaemia on prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT in normal canine plasma samples were studied using commercially available reagents and a steel ball coagulometer. Haemolysis significantly interfered with APTT (P = 0.0076) and TT (P = 0.0292). Regression analysis showed that TT was significantly shortened as haemoglobin concentrations increased. Lipaemia increased as demonstrated by regression analysis. Bilirubin significantly interfered with PT (P=0.0003) and APTT (P=0.002). Although statistically significant, none of the differences found were of clinical relevance.

Animals↗

Comparison of three prothrombin time and activated partial thromboplastin time reagent systems on the MLA 700 coagulation analyzer.

OBJECTIVE: To determine the difference in prothrombin time (PT) and activated partial thromboplastin time (aPTT) results among three reagent systems using a single analyzer instrument. DESIGN: Convenience sample of 100 patient specimens tested in duplicate with three reagent systems: Baxter-Dade, Pacific Hemostasis, and Organon Teknika. SETTING: A tertiary hospital that services other institutions within a three-state area. PATIENTS: Patients were divided into four groups: (1) normal preoperative patients who received no anticoagulants, (2) patients who received warfarin for at least the week immediately before the study, (3) patients who received heparin on the day of the testing, and (4) patients with severe liver disease accompanied by abnormal liver function tests. MAIN OUTCOME MEASURE: Coefficients of correlation of Baxter-Dade results versus the other two systems. RESULTS: PT values were significantly different in normal samples and in warfarin-treated patients. aPTT values were significantly different for normal patients and, for the Organon system only, for heparin-treated patients. When expressed as international normalized ratio (INR) values, taking reagent sensitivity into consideration, the results correlated well. Problems with precipitation when using Organon's system limited its practical utility. CONCLUSION: Compatibility between a reagent system and analyzer instrument should be verified by the manufacturer of the instrument. Use of the INR format produced more accurate and comparable results, allowing safer and more effective dosage adjustments. Laboratories should convert PT and aPTT results to the INR format routinely.

Adult↗

The European Concerted Action on Anticoagulation (ECAA) evaluation of a set of lyophilized normal plasmas to establish the normal prothrombin time for coagulometer systems.

Establishing the mean normal prothrombin time (MNPT) from fresh samples for prothrombin ratios and INR often presents difficulty in selection and collection of donors. A set of seven lyophilized normal plasmas has therefore been prepared at the ECAA Central Facility and studied at 143 laboratories in sixteen European states using coagulometers in serial field exercises. All centres tested either the high ISI ECAA rabbit or low ISI ECAA human reference thromboplastin. The MNPT of fresh plasmas and means of the lyophilized samples were closely comparable with most routine rabbit thromboplastins. Using human thromboplastins means with the lyophilized normals were marginally but significantly longer and with the bovine Thrombotest significantly shorter than the MNPT of fresh plasmas causing alterations in INR. There was no appreciable effect on INR of 2.5 and 3.5 when lyophilized normals were substituted for fresh normals with the rabbit reagents.

Animals↗

Single-dose fluconazole for vulvovaginal candidiasis: impact on prothrombin time in women taking warfarin.

OBJECTIVE: To estimate the effect of a single oral 150-mg dose of fluconazole on the prothrombin time of women on long-term warfarin therapy. METHODS: Women on warfarin therapy for 6 months or more with no change in dose within 4 weeks of the study, and a prothrombin time (PT) with an International Normalized Ratio (INR) between 2 and 3 were invited to participate. Two consecutive baseline PTs were obtained (days -1 and 0), and women were given 150 mg of fluconazole. Prothrombin times were measured on days 2, 5, and 8 of the study. The change in PT was calculated from the difference between the baseline PT on day 0 and the PT during the study period. To detect a 10% difference in a PT (approximately 2.1 seconds) of a patient with an INR of 2.0, at P < .05 and a power of 90%, 5 subjects are required. RESULTS: Six women participated. The mean (+/- standard deviation) PT for day 0 was 27.7 +/- 4.1 seconds or INR 2.6 +/- 0.4. The PT increased 11% at day 2, 34% at day 5, and 2% at day 8; these differences were not statistically significant. However, one half of the women had either a clinically relevant increase of the INR greater than 4, or bleeding that required their dosage of warfarin to be decreased. CONCLUSION: A single 150-mg oral dose of fluconazole may increase the PT to a clinically relevant level in a woman on chronic warfarin therapy. Clinicians should monitor the PT carefully after a single dose of fluconazole. LEVEL OF EVIDENCE: II-2.

Anticoagulants↗

The role of augmented Hageman factor (factor XII) titers in the cold-promoted activation of factor VII and spontaneous shortening of the prothrombin time in women using oral contraceptives.

Spontaneous shortening of the Thrombotest time and an increase in factor VII activity after cold storage of plasma in plastic vials have been described in the plasma of women using OCAs. The phenomenon if CPA is dependent on the presence of HF (factor XII) and is associated with increased plasma kallikrein activity and the disappearance of C1-INH activity. In the present study, spontaneous CPA and shortening of the prothrombin time were observed in 66% of women using OCAs. The prothrombin time was inversely related to factor VII activity and HF coagulant titer, whereas factor VII activity was strongly associated with the titer of HF. The addition of purified HF to normal plasma to a degree observed in OCA users promoted CPA generation and shortening of the prothrombin time. In contrast, reduction of the HF coagulant titer in the plasma of OCA users to a normal level prevented the rise in factor VII activity and shortening of the prothrombin time after cold storage. These data indicate that a high titer of HF, such as that observed in OCA users, is needed for the spontaneous CPA and add support to evidence linking the extrinsic and intrinsic pathways of clotting. The relationship of this in vitro phenomenon to the pathogenesis of thrombosis in women using OCAs is presently unclear. (J Lab Clin Med 99:363, 1982.)

Adult↗

Searching for inaccuracy in clinical laboratory testing using Medicare data. Evidence for prothrombin time.

OBJECTIVE: To determine if the occurrence of health outcomes following clinical laboratory testing can be used to identify types of laboratories that may be having higher than expected error rates. DESIGN: Retrospective analysis of Medicare Part B outpatient claims, Part A hospitalization bills, and death records using a case-control study. SETTING: Medicare records from six carrier territories were sampled during the period 1985 through 1987. PATIENTS: A total of 14,755 Medicare patients receiving a prothrombin time test in either a physician office laboratory or a commercial laboratory. OUTCOME MEASURES: Occurrence of a hospitalization for stroke or acute myocardial infarction, death, or no adverse outcome within 6 days of a prothrombin time. RESULTS: In physician office laboratories where prothrombin time test volume is below 40 per month, the odds that a tested patient will experience a stroke or an acute myocardial infarction are up to 1.96 and 3.43 times greater, respectively, than for a similar patient tested in a commercial laboratory. Switching from one laboratory to another between successive prothrombin time tests increased the odds of a stroke or an acute myocardial infarction by 1.57 and 1.32, respectively. Patients in two states with strong laboratory regulatory programs had fewer adverse outcomes. CONCLUSION: Examining patient outcomes subsequent to clinical laboratory testing may be a useful tool for clinical laboratory quality assurance.

Case-Control Studies↗

Recombinant activated factor VII (rFVIIa) acutely normalizes prothrombin time in patients with cirrhosis during bleeding from oesophageal varices.

BACKGROUND: Patients with cirrhosis have low levels of coagulation factors, the most pronounced deficiency being that of FVII. This may compromise haemostasis during bleeding from ruptured oesophageal varices. The objective of this trial was to evaluate the effect of rFVIIa on prothrombin time in cirrhosis patients with ongoing variceal bleeding. Safety, including signs of DIC, was monitored. METHODS: The study is a single centre, open-label trial. Ten consecutive patients with known alcoholic cirrhosis and oesophageal variceal bleeding were included. The patients received routine treatment, including Terlipressin. Each patient received one i.v. injection of rFVIIa (80 microg/kg bw). The study observation time was 12 h per patient. RESULTS: The mean age of the patients was 48 years (8 men and 2 women). The cirrhosis was classified as Child B in 5 patients and Child C in 5. At baseline, all patients had prothrombin time levels above the normal range, and all but one had FVII coagulation activity (FVII:C) levels below the normal range. rFVIIa normalized the prothrombin time in all patients within 30 min. The effect lasted for more than 4 h in 7 patients, and for about 2 h in the remaining 3 patients. Immediate bleeding control was obtained in all patients, and no patient died within the study time. There was no sign of DIC. CONCLUSIONS: rFVIIa is effective in transiently reversing the prolonged prothrombin time in cirrhosis patients with haematemesis from varices. This indicates a potential of improving haemostasis and survival in patients with compromised coagulation due to liver disease.

Adult↗

Prothrombin times and clottable fibrinogen determination on an automated coagulation laboratory (ACL-810).

Our laboratory evaluated an Automated Coagulation Laboratory (ACL-810) by Instrumentation Laboratory. Prothrombin times and fibrinogen levels from the ACL-810 were compared to results from a Fibrometer and another automated coagulation instrument - either the Coag-A-Mate (prothrombin times) or the Multistat III centrifugal analyzer (fibrinogen). The performance of different thromboplastins on the ACL-810 was also evaluated. Correlation studies revealed excellent performance characteristics and precisions for both the prothrombin time and fibrinogen not only with different instruments, but also with different thromboplastins on the ACL-810. Overall, we found the instrument to be quick, efficient, and easy to operate.

Autoanalysis↗

Comparison of anticoagulants for the preservation of prothrombin time specimens.

The value of different anticoagulants as preservatives of specimens for prothrombin time ratios is evaluated and discussed. An increase in the ratio was demonstrated with all anti-coagulants after 24 hours and this was significant when coagulometer error was taken into account. Both solid and liquid sodium citrate were superior to sodium oxalate as preservatives at 24 hours after taking off samples, but differences were slight and were not significant when coagulometer error was considered. It was concluded that for routine clinical practice, where a range of the prothrombin time ratio of 1.8/1 to 2.5/1 is allowable, there would be no significant advantage in any of the anticoagulants considered.

Anticoagulants↗

A micromethod prothrombin time for oral anticoagulant control.

A capillary prothrombin time method was described using the Australasian Reference Thromboplastin (A.R.T.). The method was used for the control of oral anticoagulant therapy in 59 patients on 225 occasions and was found to be reliable and convenient. The capillary prothrombin ratio could be correlated with the conventional plasma prothrombin ratio.

Adult↗

Preoperative screening for coagulopathy using prothrombin time and partial thromboplastin time in patients requiring primary cranial vault remodeling.

BACKGROUND: The aim of this study was to investigate the prevalence of abnormal preoperative screening prothrombin time and partial thromboplastin time in patients listed for primary cranial vault remodeling that required hematologic workup and their diagnoses and subsequent management. METHODS: This retrospective analysis was performed from January of 2000 to December of 2003 at the International Craniofacial Institute, Dallas, Texas, on a total of 168 patients. RESULTS: All patients had a normal prothrombin time. Abnormally raised partial thromboplastin time was found in six patients (prevalence of 3.57 percent), one who had factor XI deficiency, one who had a borderline factor XI deficiency and circulating inhibitor, one who had an intermittent factor XI deficiency and circulating inhibitor, one who had a borderline von Willebrand's disease with low factor XII, and the remaining two who had a circulating inhibitor of coagulation. Of these six patients, the perioperative management was altered in four of five patients, and one patient declined surgery out of fear of surgical morbidity. The surgery of one patient was aborted intraoperatively because of abnormal bleeding without clot formation after the calvarial burr holes had been drilled. The mean blood loss was 183 ml for the four patients with completed surgery and 100 ml for one patient. CONCLUSIONS: The authors conclude that even though the prevalence of abnormal screening partial thromboplastin time in these patients was low (3.57 percent), detection of an abnormal result required preoperative correction of coagulopathy in 80 percent of cases.

Blood Coagulation Disorders↗