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Thromb Haemost 1994; 71(03): 292-299
DOI: 10.1055/s-0038-1642433
Original Article
Schattauer GmbH Stuttgart

Multicentric Evaluation of a New PT Reagent Based on Recombinant Human Tissue Factor and Synthetic Phospholipids

R Bader
1   The IRCCS Maggiore Hospital and University, Policlinico Centro Hemophilia, Milano, Italy
,
P M M Mannucci
1   The IRCCS Maggiore Hospital and University, Policlinico Centro Hemophilia, Milano, Italy
,
A Tripodi
1   The IRCCS Maggiore Hospital and University, Policlinico Centro Hemophilia, Milano, Italy
,
J Hirsh
2   The Hamilton Civic Hospitals Research Centre, Hamilton, Ontario, Canada
,
F Keller
3   The Medizinische Universitätsklinik, Zentrallabor, Würzburg, Germany
,
E M Solleder
3   The Medizinische Universitätsklinik, Zentrallabor, Würzburg, Germany
,
P Hawkins
4   The Baxter Diagnostics, R & D, Miami, Florida, USA
,
M Peng
4   The Baxter Diagnostics, R & D, Miami, Florida, USA
,
H Pelzer
4   The Baxter Diagnostics, R & D, Miami, Florida, USA
,
L M Teijidor
4   The Baxter Diagnostics, R & D, Miami, Florida, USA
,
I F Ramirez
5   The Baxter Deutschland GmbH, Haemostase Europa, Unterschleißheim/München, Germany
,
H-J Kolde
5   The Baxter Deutschland GmbH, Haemostase Europa, Unterschleißheim/München, Germany
› Institutsangaben
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Publikationsverlauf

Received: 26. Oktober 1992

Accepted after revision 12. Dezember 1993

Publikationsdatum:
06. Juli 2018 (online)

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Summary

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, 11,7 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 Independent 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.

 

  • References

  • 1 Quick AJ. The prothrombin in hemophilia and in obstructive jaundice. J Biol Chem 1935; 109: 72
    PubMedSuche in Google Scholar
  • 2 Morrissey JH, Fair DS, Edgington TS. Monoclonal antibody analysis of purified and cell-associated tissue factor. Thromb Res 1988; 52: 247-61
    CrossrefPubMedSuche in Google Scholar
  • 3 ICSH/ICTH Recommendations for reporting prothrombin time in oral anticoagulant control. Loeliger EA. Thromb Haemostas 1984; 54: 155-6
    PubMedSuche in Google Scholar
  • 4 Kirkwood TB L. Calibration of reference thromboplastins and standardisation of the prothrombin time ratio. Thromb Haemostas 1983; 49: 238-44
    PubMedSuche in Google Scholar
  • 5 Loeliger EA, Poller L, Samama M, Thomson JM, Van den Besselaar AM H P, Vermylen J, Verstraete M. Questions and answers on prothrombin time standardisation in oral anticoagulant control. Thromb Haemostas 1985; 54: 515-7
    PubMedSuche in Google Scholar
  • 6 Spicer EK, Horion R, Bloem L, Bach R, Williams KR, Guha A, Kraus J, Lin TC, Nemerson Y, Königsberg WH. Isolation of cDNA coding for human tissue factor: Primary structure of the protein and cDNA. Proc Natl Acad Sei USA 1987; 84: 5148-52
    CrossrefPubMedSuche in Google Scholar
  • 7 Paborsky LR, Tate KM, Harris RJ, Yansura DG, Band L, McCray G, Gorman CM, O'Brien DP, Chang JY, Swartz JR, Fung VP, Thomas JN, Vehar GA. Purification of recombinant human tissue factor. Biochemistry 1989; 28: 8072-7
    CrossrefPubMedSuche in Google Scholar
  • 8 Kang S, Niemetz J. Purification of human brain tissue factor. Thromb Haemostas 1988; 59: 400-3
    PubMedSuche in Google Scholar
  • 9 Scarpati EM, Wen D, Broze GJ, Miletich IP, Flandermeyer RP, Siegel NR, Sadler JE. Human tissue factor: cDNA sequence and chromosome localization of the gene. Biochemistry 1987; 26: 5234-8
    CrossrefPubMedSuche in Google Scholar
  • 10 Fisher KL, Gorman CM, Gordon A, Vehar DP, O’Brien, Lawn RM. Cloning and expression of human tissue factor cDNA. Thrombosis Res 1987; 48: 89-99
    CrossrefPubMedSuche in Google Scholar
  • 11 Morrissey JH, Fakhrai H, Edgington TS. Molecular cloning of the cDNA for tissue factor, the cellular receptor for the initiation of the coagulation protease cascade. Cell 1987; 50: 129-35
    CrossrefPubMedSuche in Google Scholar
  • 12 Rehemtulla A, Pepe M, Edgington TS. High level expression of recombinant human tissue factor in Chinese hamster ovary cells as a human thromboplastin. Thromb Haemostas 1991; 65: 521-7
    PubMedSuche in Google Scholar
  • 13 Sakai T, Kisiel W. Formation of tissue factor activity following incubation of recombinant human tissue factor apoprotein with plasma lipoproteins. Thrombosis Res 1990; 60: 213-22
    CrossrefPubMedSuche in Google Scholar
  • 14 Mackman N, Morrissey JH, Fowler B, Edgington TS. Complete sequence of the human tissue factor gene, a highly regulated cellular receptor that initiates the coagulation protease cascade. Biochemistry 1989; 28: 1755-62
    CrossrefPubMedSuche in Google Scholar
  • 15 Broze GJ, Leykam JE, Schwartz BD, Miletich JP. Purification of human brain tissue factor. J Biol Chem 1985; 260: 10917-20
    PubMedSuche in Google Scholar
  • 16 Hawkins P, Tejidor LP, Estevez R, Johnson K, Murza B, Maynard J, Low-rey D, Thom R, Gaur P. Prothrombin time reagents prepared from recombinant human tissue factor produced in E. Coli. Thromb Haemostas 1991; 65: 1215
    PubMedSuche in Google Scholar
  • 17 Exner T, Rickard KA, Kronenberg H. Comparison of thromboplastins used for oral anticoagulant control. Pathology 1980; 12: 559-66
    CrossrefPubMedSuche in Google Scholar
  • 18 Hirsh J. Is the dose of warfarin prescribed by American physicians unnecessarily high?. Arch Intern Med 1987; 147: 769-71
    CrossrefPubMedSuche in Google Scholar
  • 19 Hirsh J, Levine M. Confusion over the therapeutic range for monitoring oral anticoagulant therapy in North America. Thromb Haemostas 1988; 59: 129-32
    PubMedSuche in Google Scholar
  • 20 Tejidor LM, Hawkins PA, Johnson KB, Estevez RA, Maynard JA, Gaur PK. Use of synthetic phospholipids to prepare active recombinant human tissue factor. Circulation. Supplement II 1991; 84: 598
    PubMedSuche in Google Scholar
  • 21 Spaethe R. Experience gained in the calibration of thromboplastins according to the WHO model (ISI/INR-System). Ärztl Lab 1988; 10: 293-304
    PubMedSuche in Google Scholar
  • 22 van Rijn JL M L, Schmidt NA, Rutten WP F. Correction of instrument-and reagent-based differences in determination of the international normalized ratio (INR) for monitoring anticoagulant therapy. Clin Chem 1989; 35/5: 840-3
    PubMedSuche in Google Scholar
  • 23 Poggio M, van den Besselaar AM H P, van der Velde EA, Bertina RM. The effect of some instruments for prothrombin time testing on the international sensitivity index (ISI) of two rabbit tissue thromboplastin reagents. Thromb. Haemostas 1989; 62: 868-74
    PubMedSuche in Google Scholar
  • 24 van den Besselaar AM H P, Bertina RM. Multi-center calibration of the second reference material for thromboplastin, rabbit, plain, coded CRM 149R. Thromb Haemostas 1991; 65: 263-7
    PubMedSuche in Google Scholar
  • 25 Thomson JM, Tomenson JA, Poller L. The calibration of the second primary international reference preparation for thromboplastin (thromboplastin. human, plain, coded BCT/253). Thromb Haemostas 1984; 52: 336-42
    PubMedSuche in Google Scholar
  • 26 Hawkins PL, Barrow DA, Maynard JR. A sensitive thromboplastin reagent from rabbit brain tissue factor for monitoring oral anticoagulant therapy. Thromb Haemostas 1989; 62: 530
    PubMedSuche in Google Scholar
  • 27 Eisenwiener HG, Bablok W, Bardorff W, Bender R, Markowetz D, Passing H, Spaethe R, Specht W, Volkert E. Statistische Auswertung beim Methodenvergleich. Lab med 1984; 8: 232-44
    PubMedSuche in Google Scholar
  • 28 Hell stern P, Anders CK, Oberfrank K, Faller B, Spaett A. Heparinsensitivität von Thromboplastinen zur Bestimmung der Thromboplastinzeit. Klin Lab 1992; 38: 183-8
    PubMedSuche in Google Scholar
  • 29 Brandjes DP M, Heijboer H, Büller HR, De Rijk M, Jagt H, van Wouter ten Cate J. Acenocoumarol and Heparin compared with acenocoumarol alone in the initial treatment of proximal-vein thrombosis. N Engl J Med 1992; 327: 485-9
    PubMedSuche in Google Scholar
  • 30 Poller L. When to start oral anticoagulants and the optimal range. Biol Clin Hematol 1990; 12: 113-9
    PubMedSuche in Google Scholar
  • 31 Hirsh J, Poller L, Deykin D, Levine M, Dalen JE. Optimal therapeutic range for oral anticoagulants. Chest 1989; 95: 5S-11S
    CrossrefPubMedSuche in Google Scholar
  • 32 Poller L. Therapeutic ranges for oral anticoagulation in different thromboembolic disorders. Ann Hematol 1992; 64: 52-9
    CrossrefPubMedSuche in Google Scholar
  • 33 Loeliger EA. Therapeutic target values in oral anticoagulation-Justification of Dutch policy and a warning against the so-called moderate-intensity regimens. Ann Hematol 1992; 64: 60-5
    CrossrefPubMedSuche in Google Scholar
  • 34 Turpie AG, Hirsch J, Gunstensen J. et al. Randomized comparison of two intensities of oral anticoagulation therapy after tissue valve replacement. Lancet 1988; i: 1242-5
    PubMedSuche in Google Scholar
  • 35 Hull R, Hirsh J, Jay R. et al. Different intensities of oral anticoagulant therapy in the treatment of proximal vein thrombosis. N Engl J Med 1982; 307: 1676-81
    CrossrefPubMedSuche in Google Scholar
  • 36 Millenson MM, Bauer KA, Kistler JP, Barzegar S, Tulin L, Rosenberg RD. Monitoring “mini-intensity” anticoagulation with warfarin: comparison of the prothrombin time using a sensitive thromboplastin with prothrombin fragment F1+2 levels. Blood 1992; 79: 2034-8
    PubMedSuche in Google Scholar
  • 37 Moriarty HT, Lam-Po-Tang PR L, Anastas N. Comparison of thromboplastins using the ISI and INR system. Pathology 1990; 22: 71-6
    CrossrefPubMedSuche in Google Scholar
  • 38 Morrison M, Caldwell A, McQuaker G, Fitzsimons EJ. Discrepant INR values: a comparison between Manchester and Thrombotest reagents using capillary and venous samples. Clin lab Haemat 1989; 11: 393-8
    CrossrefPubMedSuche in Google Scholar
  • 39 Gogstad GO, Wadt J, Smith P, Bryuildsrud T. Utility of a modified calibration model for reliable conversion of thromboplastin time to international normalized ratios. Thromb Haemostas 1986; 56: 178-82
    PubMedSuche in Google Scholar
  • 40 Van den Besselaar AM H P. The International Normalized Ratio for Prothrombin Time Testing in Advances in Coagulation Testing. Interpretation and Application Triplett DA. (ed), College of American Pathologists, Illinois; 1986
  • 41 Kolde HJ, Strauss J, Naumann M, Keller F, Pohl B, Zürrlein P. Evaluation of a new thromboplastin. Blut 1990; 60: 147 (abstr.).
    PubMedSuche in Google Scholar
  • 42 Sartori MT, Pontara E, Zerbinati P, Boeri G, Girolami A. Uso di una nuova thromboplastina (Thromboplastin IS) per la determinazione del tempo ti protrombina. BML 1990; 10: 31-6
    PubMedSuche in Google Scholar
  • 43 Tripodi A, Arbini A, Chantarangkul V, Mannucci PM. Recombinant tissue factor as substitute for conventional thromboplastin in the prothrombin time test. Thromb Haemostas 1992; 67: 42-5
    PubMedSuche in Google Scholar
  • 44 Hawkins P, Doobay H, Tejidor L, Johnson K, Peng M, Pelzer H, Kolde HJ, Ramirez I, Gaur P. The international normalized ratio and factor sensitivity of a prothrombin time reagent based on recombinant human tissue factor. Thromb Haemostas 1993; 65: 1215
    PubMedSuche in Google Scholar
  • 45 Vogt A, Woodhams B, Marbet GA. Erste Erfahrungen mit einem rekombi-nanten menschlichen Gewebsthromboplastin bei oraler Antikoagulation. Schweiz med Wschr 1993; 123: 74-6
    PubMedSuche in Google Scholar