Anticardiolipin Antibody Assay: a Methodological Analysis for a better Consensus in Routine Determinations

 

 Buy Article Permissions and Reprints

Summary

Despite the widely recognized practical importance of anticardiolipin (aCL) ELISA, the reliability of this test has been recently discussed. In order to investigate this area on European scale, we sent to 30 experienced centers a questionnaire focusing on the diagnostic procedures applied to patients with antiphospholipid syndrome (APS) and on the detailed protocols used to perform aCL. Anticardiolipin ELISA was found to be the most frequently performed test in patients with suspected APS, but significant difference was shown among the various protocols. The cross-laboratory multiple examination of ten serum samples evaluated independently by the 24 centers pointed out the difficulty in getting comparable results. Therefore a “consensus” protocol was derived from the aCL methods giving the best performance. The materials and reagents necessary to perform the “consensus” method, including, as putative standards, one IgG and one IgM monoclonal antibody (HCAL and EY2C9) were distributed to 19 Centers. The results of one IgG and one IgM aCL high positive sera measured in serial dilutions were compared. A progressive decrease in the variability of the values obtained for a given sample appeared evident when all the laboratories used the same standard, in their own in-house ELISA and even more in the “consensus” ELISA.

Our data show that aCL ELISA standardization is necessary in order to obtain comparable results in different laboratories.

• References

• 1 Harris EN, Gharavi AE, Boey ML, Mackworth-Young CG, Loizou S, Hughes GRV. Anticardiolipin antibodies: detection by radioimmunoassay and association with thrombosis in systemic lupus erythematosus. Lancet 1983; 2: 1211-4.
  PubMedGoogle Scholar
• 2 Wilson WA, Gharavi AE, Koike T, Lockshin MD, Branch DW, Piette JCP, Brey R, Derksen R, Harris EN, Hughes GRV, Triplett DA, Khamashta MA. International consensus statement on preliminary classification criteria for definite antiphospholipid syndrome. Arthritis Rheum 1999; 42: 1309-11.
  CrossrefPubMedGoogle Scholar
• 3 Brandt JT, Triplett DA, Alving B, Scharrer I. Criteria for the diagnosis of lupus anticoagulants: an update. Thromb Haemost 1995; 74: 1185-90.
  Article in Thieme ConnectPubMedGoogle Scholar
• 4 Triplett DA. Many faces of lupus anticoagulant. Lupus 1998; 7 S2 18-22.
  CrossrefPubMedGoogle Scholar
• 5 Harris EN, Gharavi AE, Patel SP, Hughes GRV. Evaluation of the anticardiolipin antibody test: Report of a standardization workshop held in April 1986. Clin Exp Immunol 1987; 68: 215-22.
  PubMedGoogle Scholar
• 6 Harris EN, Gharavi AE, Patel SP, Hughes GRV. Anticardiolipin antibody testing. The need for standardization. Arthritis Rheum 1987; 30: 835-6.
  CrossrefPubMedGoogle Scholar
• 7 Harris EN. The Second International Anti-Cardiolipin Standardization Workshop/The Kingstone Anti-Phospholipid Antibody Study (KAPS) Group. Am J Clin Pathol 1990; 94: 476-84.
  CrossrefPubMedGoogle Scholar
• 8 Harris EN, Pierangeli S, Birch D. Anticardiolipin Wet Workshop Report. Fifth International Symposium on Antiphospholipid antibodies. Am J Clin Pathol 1994; 101: 616-24.
  CrossrefPubMedGoogle Scholar
• 9 Kalden JR. WHO/IUIS Standardization Programme. Clin Exp Rheumatol 1992; 10 (Suppl. 05) 513-4.
  PubMedGoogle Scholar
• 10 Coulam CB, McIntyre JA, Wagenknecht D, Rote N. Interlaboratories inconsistencies in detection of anticardiolipin antibodies. Lancet 1990; 335 (suppl. 8693): 865.
  PubMedGoogle Scholar
• 11 Peaceman AM, Silver RK, Mac Gregor SN, Socol ML. Interlaboratory variation in antiphospholipid antibody testing. Am J Obstet Gynecol 1992; 166: 1780-7.
  CrossrefPubMedGoogle Scholar
• 12 Reber G, Arvieux J, Comby E. Multicenter evaluation of nine kits for the quantitation of anticardiolipin antibodies. Thromb Haemost 1995; 52: 444-52.
  Article in Thieme ConnectPubMedGoogle Scholar
• 13 Favaloro EJ, Silvestrini R, Mohammed A. Clinical utility of anticardiolipin antibody assays: high inter-laboratory variation and limited consensus by participants of external quality assurance program signals a cautious approach. Pathology 1999; 31 (Suppl. 02) 142-7.
  CrossrefPubMedGoogle Scholar
• 14 Ginsberg JS, Wells PS, Brill-Edwards P, Donovan D, Moffat K, Johnston M, Stevens P, Hirsh J. Antiphospholipid antibodies and venous thromboembolism. Blood 1995; 86 (10) 3685-91.
  PubMedGoogle Scholar
• 15 Ichikawa K, Tsutsumi A, Atsumi T, Matsuura E, Kobayashi S, Hughes GRV, Khamashta M, Koike T. A chimeric antibody with the human γ1 constant region as a putative standard for assays to detect IgG β-2-glycoprotein I antibodies. Arthritis Rheum 1999; 42: 2461-70.
  CrossrefPubMedGoogle Scholar
• 16 Ichikawa K, Khamashta M, Koike T, Matsuura E, Hughes GRV. β-2-glyco-protein I reactivity of monoclonal anticardiolipin antibodies from patients with the antiphospholipid syndrome. Arthritis Rheum 1994; 37: 1453-61.
  CrossrefPubMedGoogle Scholar
• 17 Nilsson IM, Astedt B, Hedner U. Intrauterine death and circulating anticoagulant “antithromboplastin”. Acta Med Scand 1975; 197: 153-9.
  PubMedGoogle Scholar
• 18 Bowie EJ, Thompson Jr JH, Pascuzzi Jr CA. Thrombosis in systemic lupus erythematosus despite circulating anticoagulants. J Lab Clin Med 1963; 62: 416-30.
  PubMedGoogle Scholar
• 19 Alarcon-Segovia D, Osmundson PJ. Peripheral vascular syndromes associated with systemic lupus erythematosus. Ann Intern Med 1965; 62: 416-30.
  PubMedGoogle Scholar
• 20 Alagille D, Crosnier J, Soulier JP. Anticoagulant circulant à activité anti-thromboplastique. Rev Franç et Clin Biol 1956; 1: 335-45.
  PubMedGoogle Scholar
• 21 Soulier JP, Boffa MC. Avortements à repetition, thromboses et anticoagulant circulant antithromboplastine. Nouv Presse Med 1980; 9: 859-64.
  PubMedGoogle Scholar
• 22 Koike T, Sueishi M, Funaki H, Tomioka H, Yoshida S. Anti-phospholipid antibodies and biological false positive serological test for syphilis in patients with systemic lupus erythematosus. Clin Exp Immunol 1984; 56: 193-9.
  PubMedGoogle Scholar
• 23 Asherson RA, Khamashta MA, Ordi-Ros J. The “primary” antiphospholipid syndrome: Major clinical and serological features. Medicine 1989; 68: 366-74.
  CrossrefPubMedGoogle Scholar
• 24 Alarcon-Segovia D, Sanchez-Guerrero J. Primary antiphospholipid syndrome. J Rheumatol 1989; 16: 482-4.
  PubMedGoogle Scholar
• 25 Roubey RAS. Antiphospholipid antibody syndrome. In: Arthritis and allied conditions: a textbook of rheumatology. Koopman WJ. (ed) Philadelphia: Lippincott, Williams & Wilkins; 1997: 1393-406.
  Google Scholar
• 26 Asherson RA, Cervera R. The Antiphospholipid Syndrome. In: Textbook of Rheumatology. Kelly WN, Ruddy S, Harris ED, Sledge CB. (eds). Philadelphia: WB Saunders Company; 1997: 1057-64.
  Google Scholar
• 27 Harris EN. Antiphospholipid Syndrome. In: Rheumatology. Klippel JH, Dieppe PA. (eds). London: Mosby; 1998. sect. 7, 35.1-6.
  Google Scholar
• 28 Tincani A, Spatola L, Cattaneo R, Balestrieri G. Antiphospholipid ELISA: The state of the art. Lupus 1996; 5: 477-9.
  CrossrefPubMedGoogle Scholar
• 29 Tincani A, Brey R, Balestrieri G, Vitali C, Doria A, Galeazzi M, Meroni PL, Migliorini P, Neri R, Tavoni A, Bombardieri S. International survey on the management of patients with SLE. II. The result of a questionnaire regarding neuropsychiatric manifestations. Clin Exp Rheumatol 1996; 14 (Suppl. 16) 23-9.
  PubMedGoogle Scholar
• 30 Harris EN, Chan JKH, Asherson RA, Aber VR, Gharavi AE, Hughes GRV. Thrombosis, recurrent fetal loss and thrombocytopenia. Predictive value of the anticardiolipin antibody test. Arch Intern Med 1986; 146: 2153-6.
  CrossrefPubMedGoogle Scholar
• 31 Tan EM, Smolen JS, McDougal JS, Butcher BT, Chonn D, Dawkins R, Fritzler MJ, Gordon T, Hardin JA, Kalden JR, Lahita RG, Maini RN, Roth-field NF, Smeenk R, Takasaki Y, van Venrooij WJ, Week A, Wilson M, Koziol JA. A critical evaluation of enzyme immunoassays for detection of antinuclear autoantibodies of defined specificities. Arthritis Rheum 1999; 42: 455-64.
  CrossrefPubMedGoogle Scholar