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Thromb Haemost 1993; 70(06): 0925-0931
DOI: 10.1055/s-0038-1649701
Original Article
Clinical Studies
Schattauer GmbH Stuttgart

The Textarin/Ecarin Ratio: A Confirmatory Test for Lupus Anticoagulants

Douglas A Triplett
The Department of Research, Ball Memorial Hospital, Muncie, Indiana, USA, and Pentapharm AG, Basel, Switzerland
,
Kurt F Stocker
The Department of Research, Ball Memorial Hospital, Muncie, Indiana, USA, and Pentapharm AG, Basel, Switzerland
,
Gail A Unger
The Department of Research, Ball Memorial Hospital, Muncie, Indiana, USA, and Pentapharm AG, Basel, Switzerland
,
Linda K Barna
The Department of Research, Ball Memorial Hospital, Muncie, Indiana, USA, and Pentapharm AG, Basel, Switzerland
› Author Affiliations
Further Information

Publication History

Received 14 May 1993

Accepted after revision 11 August 1993

Publication Date:
06 July 2018 (online)

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Summary

Lupus anticoagulants (LA) are immunoglobulins (IgG, IgM, IgA or a mixture) which interfere with in vitro phospholipid (PL) dependent tests of coagulation (e.g. APTT, KCT, dilute Russell Viper Venom Time). LA are heterogeneous; consequently, the laboratory diagnosis is difficult and relies on multiple tests. We have developed a sensitive and relatively specific confirmatory test system based on fractions of two snake venoms. Textarin®, a protein fraction of Pseudonaja textilis venom (Australian Eastern brown snake), activates prothrombin in the presence of PL, factor V and calcium ions. Ecarin, a protein fraction of Echis carinatus venom, will activate prothrombin in the absence of any cofactors. The activation of prothrombin by Textarin yields thrombin while Ecarin yields meizothrombin. In the presence of LA, the Textarin time is prolonged and the Ecarin time is unaffected. The test results are reported as a ratio of Textarin/Ecarin times (abnormal greater than 1.3). We have evaluated this test system in the following patient populations: LA positive, therapeutically heparinized, stable oral anticoagulated, liver disease, routine preoperative, anticardiolipin antibody positive LA negative, hemophilia A, various other hereditary factor deficiencies or dysfunctional proteins, and specific inhibitors of factor V and factor VTII. The LA positive patients represented a mixed population of autoimmune disease, drug-induced and post-infectious states. Our findings indicate the sensitivity of the Textarin/Ecarin system in the confirmation of LA. In order to use the test system most effectively, it is recommended to incorporate poly-brene with Textarin when evaluating heparinized samples. Factor V deficiency and specific inhibitors of factor V yielded, in some instances, false positive results.

 

  • References

  • 1 Triplett DA, Brandt JT. Lupus anticoagulants: misnomer, paradox, riddle, epiphenomenon. Hematol Pathol 1988; 2: 121-143
    PubMedGoogle Scholar
  • 2 Love PE, Santoro SA. Antiphospholipid antibodies: anticardiolipin and the lupus anticoagulant in systemic lupus erythematosus (SLE) and in non-SLE disorders. Ann Int Med 1990; 112: 682-698
    CrossrefPubMedGoogle Scholar
  • 3 McNeil HP, Chesterman CN, Krilis SA. Immunology and clinical importance of antiphospholipid antibodies. Adv Immunol 1991; 49: 193-280
    PubMedGoogle Scholar
  • 4 Triplett DA. Antiphospholipid antibodies and recurrent pregnancy loss. Am J Reprod Immunol 1989; 20: 52-67
    CrossrefPubMedGoogle Scholar
  • 5 Green D, Hougie C, Kazmier FJ, Lechner K, Mannucci PM, Rizza CR, Sultan Y. Report of the working party on acquired inhibitors of coagulation: studies of the “lupus anticoagulant”. Thromb Haemost 1983; 49: 144-146
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Triplett DA, Brandt JT, Musgrave KA, Orr CA. The relationship between lupus anticoagulants and antibodies to phospholipid. JAMA 1988; 259: 550-554
    CrossrefPubMedGoogle Scholar
  • 7 Asherson RA, Khamashta MA, Ordi-Ros J, Derksen RH, Machin SJ, Barquinero J, Outt HH, Harris EN, Vilardell-Torres M, Hughes GR. The “primary” antiphospholipid syndrome: major clinical and serological features. Medicine 1989; 68: 366-374
    CrossrefPubMedGoogle Scholar
  • 8 Mackworth-Young CG. Primary antiphospholipid syndrome: features of patients with raised anticardiolipin antibodies and no other disorder. An Rheum Dis 1989; 48: 362-367
    CrossrefPubMedGoogle Scholar
  • 9 Triplett DA. Coagulation assays for the lupus anticoagulant: review and critique of current methodology. Stroke 1992; 23 (Suppl. 01) 111-114
    PubMedGoogle Scholar
  • 10 Triplett DA, Brandt JT, Maas RL. The laboratory heterogeneity of lupus anticoagulants. Arch Pathol Lab Med 1985; 109: 946-951
    PubMedGoogle Scholar
  • 11 Exner T, Triplett DA, Taberner D, Machin SJ. Guidelines for testing and revised criteria for lupus anticoagulants. SSC Subcommittee for the Standardization of Lupus Anticoagulants. Thromb Haemost 1991; 65: 320-322
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Schleider MA, Nachman RL, Jaffe EA, Coleman M. A clinical study of the lupus anticoagulant. Blood 1976; 48: 499-509
    PubMedGoogle Scholar
  • 13 Thiagarajan P, Pengo V, Shapiro SS. The use of the dilute Russell viper venom time for the diagnosis of lupus anticoagulants. Blood 1986; 68: 869-874
    CrossrefPubMedGoogle Scholar
  • 14 Alving BM, Barr CF, Johansen LE, Tang DB. Comparison between a one-point dilute phospholipid APTT and the dilute Russell viper venom time for verification of lupus anticoagulants. Thromb Haemost 1992; 67: 672-678
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Triplett DA, Brandt JT, Kaczor D, Schaeffer J. Laboratory diagnosis of lupus inhibition: a comparison of the tissue thromboplastin inhibition procedure with a new platelet neutralization procedure. Am J Clin Pathol 1983; 79: 678-682
    CrossrefPubMedGoogle Scholar
  • 16 Rosove MH, Ismail M, Kozoil BJ, Runge A, Kasper CK. Lupus anticoagulants: improved diagnosis with a kaolin clotting time using rabbit brain phospholipid in standard and high concentrations. Blood 1986; 68: 472-478
    PubMedGoogle Scholar
  • 17 Amout J, Huybrechts E, Vanrusselt E, Vermylen J. A new lupus anticoagulant neutralization test based on platelet-derived vesicles. Br J Haematol 1992; 80: 341-346
    CrossrefPubMedGoogle Scholar
  • 18 Rauch J, Tannenbaum M, Janoff AS. Distinguishing plasma lupus anticoagulants from anti-factor antibodies using hexagonal (II) phase phospholipids. Thromb Haemost 1989; 62: 892-896
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Rosing J, Zwaal RFA, Tans G. Snake venom prothrombin activators. In: Hemostasis and Animal Venoms. Pirkle H, Markland Jr FS. (eds) New York, Basel: Marcel Dekker, Inc; 1988: 03-27
    Google Scholar
  • 20 Speijer H, Govers-Riemslalg JW, Zwaal RF, Rosing J. Prothrombin activation by an activator from the venom of Oxyuranus scutellatus (Taipan snake). J Biol Chem 1986; 260: 9366-9372
    PubMedGoogle Scholar
  • 21 Morita T, Iwanaga S, Suzuki T. Activation of bovine prothrombin by an activator isolated from Echis carinatus venom . Thromb Res 1976; 8: 59-76
    PubMedGoogle Scholar
  • 22 Triplett DA, Harms CS. Procedures for the coagulation laboratory. Am Soc Clin Pathol. Chicago, Illinois: 1981
    Google Scholar
  • 23 Masci PP, Whitaker AN, deJersey J. Purification and characterization of a prothrombin activator from the venom of the Australian brown snake Pseudonaja textilis . Biochem Int 1988; 17: 825-835
    PubMedGoogle Scholar
  • 24 Pirkle H, Marsh H. Nomenclature of exogenous haemostatic factors. Thromb Haemost 1991; 66: 264
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Bell WN, Alton HB. A brain extract as a substitute for platelet suspensions in the thromboplastin generation test. Nature Lond 1954; 174: 880-881
    CrossrefPubMedGoogle Scholar
  • 26 Miletich JP, Majerus DW, Majerus PW. Patients with congenital factor V deficiency have decreased factor Xa binding sites on their platelets. J Clin Invest 1978; 62: 824-831
    CrossrefPubMedGoogle Scholar
  • 27 Triplett DA. Screening for the lupus anticoagulant. Res Clin Lab 1989; 19: 379-389
    PubMedGoogle Scholar
  • 28 Harris EN, Gharavi AE, Patel SA, Hughes GRV. Evaluation of the anticardiolipin test: report of an international workshop held 4 April 1986. Clin Exp Immunol 1987; 68: 215-222
    PubMedGoogle Scholar
  • 29 National Committee for Clinical Laboratory Standards. Collection, transport, and processing of blood specimens for coagulation testing and performance of coagulation assays. Approved Guidelines. NCCLS Document H21-A2 2nd. edition Villanova, Pennsylvania: NCCLS; 1991
    Google Scholar
  • 30 Mannucci PM, Canciani MT, Mari D, Meucci P. The varied sensitivity of partial thromboplastin and prothrombin time reagents in the demonstration of lupus-like anticoagulant. Scand J Haematol 1979; 22: 423-432
    PubMedGoogle Scholar
  • 31 Rhee M, Morris S, Kosow DP. Role of meizothrombin and meizothrombin-(des FI) in the conversion of prothrombin to thrombin by the Echis carinatus venom coagulant. Biochemistry 1982; 21: 3437-3443
    CrossrefPubMedGoogle Scholar
  • 32 Govers-Riemslag JWP, Speijer H, Zwaal RFA, Rosing J. Purification and characterization of the prothrombin activator from Oxyuranus scutellatus (Taipan snake). In: Hemostasis and Animal Venoms. Pirkle H, Markland Jr FS. (eds) New York, Basel: Marcel Dekker, Inc; 1988: 41-53
    Google Scholar
  • 33 Triplett DA, Hannah D, Barna L. Hexagonal (II) phase phospholipid neutralization of lupus anticoagulants: a specific and sensitive confirmatory procedure. Thromb Haemost 1991; 65: 1255 (abstract 2021)
    PubMedGoogle Scholar
  • 34 Oosting JD, Derksen RHWM, Entjes HTI, Bouma BN, deGroot PG. Lupus anticoagulant activity is frequently dependent on the presence of B2Glycoprotein I. Thromb Haemost 1992; 67: 499-502
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Bevers EM, Galli M, Barbui T, Comfurius P, Zwaal RFA. Lupus anticoagulant IGG’s (LA) are not directed to phospholipid only but to a complex of lipid bound prothrombin. Thromb Haemost 1991; 66: 629-632
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Vermylen J, Arnout J. Is the antiphospholipid syndrome caused by antibodies directed against physiologically relevant phospholipidprotein complexes?. J Lab Clin Med 1992; 120: 10-12
    PubMedGoogle Scholar
  • 37 Oosting JD, Derksen RHWM, Bobbink IWG, Hackeng TM, Bouma BN, deGroot PG. Antiphospholipid antibodies directed against a combination of phospholipids with prothrombin, protein C or protein S: an explanation for their pathogenic mechanism?. Blood 1993; 81: 2618-2625
    CrossrefPubMedGoogle Scholar
  • 38 Zehnder JL, Leung LLK. Development of antibodies to thrombin and factor V with recurrent bleeding in a patient exposed to topical bovine thrombin. Blood 1990; 76: 2011-2016
    PubMedGoogle Scholar
  • 39 Nichols WL, Daniels TM, Fisher PK, Owen WG, Figueroa PI, Pineda AA. Inhibitors of coagulation factor V and thrombin associated with surgical use of topical bovine thrombin or fibrin “glue”. Blood 1991; 78 (Suppl. 01) 63 a (abstract 241)
    PubMedGoogle Scholar
  • 40 Spero JA, Triplett DA, Cmolik BL, Reid C, Clark RE. A family of clotting factor inhibitors primarily involving FV and FXI occurring after cardiovascular and neurosurgical procedures. Blood 1991; 78 (Suppl. 01) 63 a (abstract 243)
    PubMedGoogle Scholar
  • 41 Rapaport SI, Zivelin A, Minow RA, Hunter CS, Donnelly K. Clinical significance of antibodies to bovine and human thrombin and factor V after surgical use of bovine thrombin. Am J Clin Pathol 1992; 97: 84-91
    CrossrefPubMedGoogle Scholar