Detection of Lupus-Like Anticoagulants by an Enzyme-Linked Immunosorbent Assay Using a Partial Thromboplastin as Antigen; A Comparative Study^[*]

Summary

Clotting assays allow qualitative rather than quantitative detection of the lupus anticoagulant. We have therefore studied the usefulness of an ELISA using a commercial partial thromboplastin, Thrombofax, oS antigen; the results obtained on 146 selected patient plasmas were compared to the results of coagulation tests (kaolin clotting time, tissue thromboplastin inhibition test, activated partial thromboplastin time) and of ELISAs using cardiolipin or phosphatidylserine as antigen. While satisfactory agreement was found within the group of coagulation tests or that of ELISAs, only a moderate agreement was obtained between clotting tests and ELISAs, the best being with the partial thromboplastin ELISA using low plasma dilutions. The study further indicates that ELISA techniques cannot entirely replace coagulation tests for the detection of a lupus anticoagulant, even when a partial thromboplastin is used as antigen. On the other hand, coagulation tests are less sensitive than ELISAs for the detection of antiphospholipid antibodies.

Dedicated to Professor M. Verstraete on the occasion of his 65th birthday

• References

• 1 Shapiro SS, Thiagarajan P. Lupus anticoagulants. Progr Hemostas Thromb 1982; 6: 263-285
• 2 Thiagarajan P, Shapiro SS, De Marco L. Monoclonal immunoglobulin MA, coagulation inhibitor with phospholipid specificity. Mechanism of a lupus anticoagulant. J Clin Invest 1980; 66: 397-405
• 3 Feinstein DI, Rapaport SI. Acquired inhibitors of blood coagulation. Progr Hemostas Thromb 1972; 1: 75-95
• 4 Conley CL, Hartmann RC. A hemorrhagic disorder caused by circulating anticoagulant in patients with disseminated lupus erythematosus. J Clin Invest 1952; 31: 621-622
• 5 Vermylen J, Blockmans D, Spitz B, Deckmyn H. Thrombosis and immune disorders. Clin Haematol 1986; 15: 393-412
• 6 Lechner K. Acquired inhibitors in non hemophiliac patients. Haemostasis 1974; 3: 65-93
• 7 Lechner K. Lupus anticoagulants and thrombosis. In: Thrombosis and Haemostasis, 1987 Verstraete M, Vermylen J, Lijnen HR, Amout J. (eds). Leuven Univ Press; Leuven: 1987: 525-547
• 8 Margolius A, Jackson DP, Ratnoff OD. Circulating anticoagulants: a study of 40 cases and a review of the literature. Medicine 1961; 40: 145-202
• 9 Gastineau DA, Kazmier FJ, Nichols WL, Bowie EJ W. Lupus anticoagulant: an analysis of the clinical and laboratory features of 219 cases. Am J Hematol 1985; 19: 265-275
• 10 Exner T, Rickard KA, Kronenberg H. A sensitive test demonstrating lupus anticoagulant and its behavioural patterns. Br J Haematol 1978; 40: 143-151
• 11 Rosner E, Pauzner R, Lusky A, Modan M, Many A. Detection and quantitative evaluation of lupus circulating anticoagulant activity. Thromb Haemostas 1987; 57: 144-147
• 12 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
• 13 Schleider MA, Nachman RL, Jaffe EA, Coleman M. A clinical study of the lupus anticoagulant. Blood 1976; 48: 499-510
• 14 Mannucci PM, Canciani MT, Mari D, Meucci P. The varied sensitivity of partial thromboplastin time reagents in the demonstration of the lupus-like anticoagulant. Scand J Haematol 1979; 22: 423-432
• 15 Triplett DA, Brandt JT, Kaczor D, Schaeffer J. Laboratory diagnosis of lupus inhibitors: a comparison of the tissue thromboplastin inhibition procedure with a new platelet neutralization procedure. Am J Clin Pathol 1983; 79: 678-682
• 16 Rosove MH, Ismail M, Koziol 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
• 17 Loizou S, McCrea JD, Rudge AC, Reynolds R, Boyle CC, Harris EN. Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results. Clin Exp Immunol 1985; 62: 738-745
• 18 Branch DW, Rote AC, Dostal DA, Scott JR. Association of lupus anticoagulant with antibody against phosphatidylserine. Clin Immunol Immunopathol 1987; 42: 63-75
• 19 Branch DW, Rote NS, Scott JR. The demonstration of lupus anticoagulant by an enzyme-linked immunosorbent assay. Clin Immunol Immunopathol 1986; 39: 298-307
• 20 Vermylen J, Amout J, Debois P, Falcon C, Huybrechts E, Vanrusselt M. An enzyme-linked immunosorbent assay for antiphospholipid antibodies using Thrombofax. Clin Exp Rheumatol 1988; 6: 214
• 21 Lopez-Soto A, Casals FJ, Font J, Cervera R, Ingelmo M. Detection of antiphospholipid antibodies by ELISA-method with bovine thromboplastin as antigen. Thromb Res 1989; 53: 623-628
• 22 Petri M, Rheinschmidt M, Whiting-O’Keefe Q, Hellmann D, Corash L. The frequency of lupus anticoagulant in systemic lupus erythematosus. Ann Intern Med 1987; 106: 524-531
• 23 Triplett DA, Brandt JT, Musgrave K, Orr CA. The relationship between lupus anticoagulants and antibodies to phospholipids. JAMA 1988; 259: 550-554
• 24 Snedecor GW, Cochran WG. Statistical Methods. The Iowa State University Press; Amer, Iowa: 1967: 186
• 25 Kitchens CS. Prolonged activated partial thromboplastin time of unknown etiology: a prospective study of 100 consecutive cases referred for consultation. Am J Hematol 1988; 27: 38-45
• 26 Bonnar J. The blood coagulation and fibrinolytic system during pregnancy. Clin Obstet Gynaecol 1975; 2: 321-325
• 27 Stirling Y, Woolf L, North WR S, Seghatchian MJ, Meade TW. Haemostasis in normal pregnancy. Thromb Haemostas 1984; 52: 176-182
• 28 Staub HL, Harris EN, Khamashta MA, Savidge G, Chahade WH, Hughes GR. Antibody to phosphatidylethanolamine in a patient with lupus anticoagulant and thrombosis. Ann Rheum Dis 1989; 48: 166-169