Interference in Antiphospholipid Antibody Assays

 

 Buy Article Permissions and Reprints

Abstract

Antiphospholipid antibodies (aPL) are detected with two types of laboratory tests: first, antigen-specific immunoassays for the determination of antibodies against cardiolipin, β₂-glycoprotein I and other phospholipids, and phospholipid−protein complexes; second, functional (coagulation) assays for the detection of lupus anticoagulants. Both aPL immunoassays and coagulation assays are prone to interferences, and clinicians need to be aware of the limitations of these assays. Interference is a clinically significant bias in the measured analyte concentration due to the effect of another component or property of the sample. Besides immune-mediated interferences (such as heterophile or human anti-animal antibodies, rheumatoid factor, high immunoglobulin levels, or factor inhibitors), aPL assays are uniquely affected by anticoagulants and the presence of residual platelets in test plasma. Interferences are usually analyte- and assay-specific and may go unrecognized in routine laboratory practice. Despite advances in our knowledge on the mechanisms of interferences in aPL assays, it is unlikely that total elimination will be possible.

• References

• 1 Clinical and Laboratory Standards Institute (CLSI). Interference Testing in Clinical Chemistry; Approved Guideline–Second Edition. CLSI document EP7–A2. Wayne, PA: Clinical and Laboratory Standards Institute; 2005
  Google Scholar
• 2 Ismail AA, Walker PL, Cawood ML, Barth JH. Interference in immunoassay is an underestimated problem. Ann Clin Biochem 2002; 39 (Pt 4) 366-373
  CrossrefPubMedGoogle Scholar
• 3 Cole LA, Rinne KM, Shahabi S, Omrani A. False-positive hCG assay results leading to unnecessary surgery and chemotherapy and needless occurrences of diabetes and coma. Clin Chem 1999; 45 (2) 313-314
  PubMedGoogle Scholar
• 4 Després N, Grant AM. Antibody interference in thyroid assays: a potential for clinical misinformation. Clin Chem 1998; 44 (3) 440-454
  PubMedGoogle Scholar
• 5 Miyakis S, Lockshin MD, Atsumi T , et al. International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome (APS). J Thromb Haemost 2006; 4 (2) 295-306
  CrossrefPubMedGoogle Scholar
• 6 Clinical and Laboratory Standards Institute (CLSI). Immunoassay Interference by Endogeneous Antibodies; Approved Guideline. CLSI document I/LA30-A. Clinical and Laboratory Standards Institute: Wayne, PA; 2008
  Google Scholar
• 7 Kano K, Merrick JM, Milgrom F. Classification of human heterophile antibodies. Int Arch Allergy Appl Immunol 1984; 73 (4) 373-377
  CrossrefPubMedGoogle Scholar
• 8 Boscato LM, Stuart MC. Incidence and specificity of interference in two-site immunoassays. Clin Chem 1986; 32 (8) 1491-1495
  PubMedGoogle Scholar
• 9 Patarca R, Fletcher MA. Structure and pathophysiology of the erythrocyte membrane-associated Paul-Bunnell heterophile antibody determinant in Epstein-Barr virus-associated disease. Crit Rev Oncog 1995; 6 (3–6) 305-326
  CrossrefPubMedGoogle Scholar
• 10 Tate J, Ward G. Interferences in immunoassay. Clin Biochem Rev 2004; 25 (2) 105-120
  PubMedGoogle Scholar
• 11 Preissner CM, O'Kane DJ, Singh RJ, Morris JC, Grebe SK. Phantoms in the assay tube: heterophile antibody interferences in serum thyroglobulin assays. J Clin Endocrinol Metab 2003; 88 (7) 3069-3074
  CrossrefPubMedGoogle Scholar
• 12 Willman JH, Martins TB, Jaskowski TD, Hill HR, Litwin CM. Heterophile antibodies to bovine and caprine proteins causing false-positive human immunodeficiency virus type 1 and other enzyme-linked immunosorbent assay results. Clin Diagn Lab Immunol 1999; 6 (4) 615-616
  PubMedGoogle Scholar
• 13 Spencer DV, Nolte FS, Zhu Y. Heterophilic antibody interference causing false-positive rapid human immunodeficiency virus antibody testing. Clin Chim Acta 2009; 399 (1–2) 121-122
  CrossrefPubMedGoogle Scholar
• 14 Levinson SS, Miller JJ. Towards a better understanding of heterophile (and the like) antibody interference with modern immunoassays. Clin Chim Acta 2002; 325 (1–2) 1-15
  CrossrefPubMedGoogle Scholar
• 15 Kricka LJ. Human anti-animal antibody interferences in immunological assays. Clin Chem 1999; 45 (7) 942-956
  CrossrefPubMedGoogle Scholar
• 16 Klee GG. Human anti-mouse antibodies. Arch Pathol Lab Med 2000; 124 (6) 921-923
  PubMedGoogle Scholar
• 17 Kricka LJ, Schmerfeld-Pruss D, Senior M, Goodman DB, Kaladas P. Interference by human anti-mouse antibody in two-site immunoassays. Clin Chem 1990; 36 (6) 892-894
  PubMedGoogle Scholar
• 18 Rothberg RM, Farr RS. Anti-bovine serum albumin and anti-alpha lactalbumin in the serum of children and adults. Pediatrics 1965; 35: 571-588
  PubMedGoogle Scholar
• 19 Berth M, Bosmans E, Everaert J , et al. Rheumatoid factor interference in the determination of carbohydrate antigen 19-9 (CA 19-9). Clin Chem Lab Med 2006; 44 (9) 1137-1139
  CrossrefPubMedGoogle Scholar
• 20 Barceló Martín B, Marquet P, Ferrer JM , et al. Rheumatoid factor interference in a tacrolimus immunoassay. Ther Drug Monit 2009; 31 (6) 743-745
  PubMedGoogle Scholar
• 21 Norden AG, Jackson RA, Norden LE, Griffin AJ, Barnes MA, Little JA. Misleading results from immunoassays of serum free thyroxine in the presence of rheumatoid factor. Clin Chem 1997; 43 (6 Pt 1) 957-962
  PubMedGoogle Scholar
• 22 Shmerling RH, Delbanco TL. The rheumatoid factor: an analysis of clinical utility. Am J Med 1991; 91 (5) 528-534
  CrossrefPubMedGoogle Scholar
• 23 Stevenson DL, Harris AG, Neal KR, Irving WL ; Trent HCV Study Group. The presence of rheumatoid factor in sera from anti-HCV positive blood donors interferes with the detection of HCV-specific IgM. J Hepatol 1996; 25 (5) 621-626
  CrossrefPubMedGoogle Scholar
• 24 Meurman OH, Ziola BR. IgM-class rheumatoid factor interference in the solid-phase radioimmunoassay of rubella-specific IgM antibodies. J Clin Pathol 1978; 31 (5) 483-487
  CrossrefPubMedGoogle Scholar
• 25 Champsaur H, Fattal-German M, Arranhado R. Sensitivity and specificity of viral immunoglobulin M determination by indirect enzyme-linked immunosorbent assay. J Clin Microbiol 1988; 26 (2) 328-332
  PubMedGoogle Scholar
• 26 Wong RC, Gillis D, Adelstein S , et al. Consensus guidelines on anti-cardiolipin antibody testing and reporting. Pathology 2004; 36 (1) 63-68
  CrossrefPubMedGoogle Scholar
• 27 Wong RC, Favaloro EJ, Adelstein S , et al. Consensus guidelines on anti-β 2 glycoprotein I testing and reporting. Pathology 2008; 40 (1) 58-63
  CrossrefPubMedGoogle Scholar
• 28 Pierangeli SS, Harris EN. A protocol for determination of anticardiolipin antibodies by ELISA. Nat Protoc 2008; 3 (5) 840-848
  CrossrefPubMedGoogle Scholar
• 29 Tincani A, Allegri F, Balestrieri G , et al. Minimal requirements for antiphospholipid antibodies ELISAs proposed by the European Forum on antiphospholipid antibodies. Thromb Res 2004; 114 (5–6) 553-558
  CrossrefPubMedGoogle Scholar
• 30 Reber G, Tincani A, Sanmarco M, de Moerloose P, Boffa MC ; Standardization group of the European Forum on Antiphospholipid Antibodies. Proposals for the measurement of anti-beta2-glycoprotein I antibodies. Standardization group of the European Forum on Antiphospholipid Antibodies. J Thromb Haemost 2004; 2: 1860-1862
  CrossrefPubMedGoogle Scholar
• 31 Galperin I, Fortin PR, Subang R, Newkirk MM, Rauch J. A subset of rheumatoid factors crossreacts with cardiolipin in patients positive for IgM rheumatoid factor and anticardiolipin antibodies. J Rheumatol 2000; 27 (3) 820-821
  PubMedGoogle Scholar
• 32 Agopian MS, Boctor FN, Peter JB. False-positive test result for IgM anticardiolipin antibody due to IgM rheumatoid factor. Arthritis Rheum 1988; 31 (9) 1212-1213
  CrossrefPubMedGoogle Scholar
• 33 Lakos G, Teodorescu M. IgM, but not IgA rheumatoid factor interferes with anti-cardiolipin and antiβ2 glycoprotein I measurements: a quantitative analysis. Lupus 2011; 20 (6) 614-619
  CrossrefPubMedGoogle Scholar
• 34 Villalta D, Crovatto M, Stella S, Tonutti E, Tozzoli R, Bizzaro N. False positive reactions for IgA and IgG anti-tissue transglutaminase antibodies in liver cirrhosis are common and method-dependent. Clin Chim Acta 2005; 356 (1–2) 102-109
  CrossrefPubMedGoogle Scholar
• 35 Bizzaro N, Tampoia M, Villalta D , et al. Low specificity of anti-tissue transglutaminase antibodies in patients with primary biliary cirrhosis. J Clin Lab Anal 2006; 20 (5) 184-189
  CrossrefPubMedGoogle Scholar
• 36 Vermeersch P, Coenen D, Geboes K, Mariën G, Hiele M, Bossuyt X. Use of likelihood ratios improves clinical interpretation of IgA anti-tTG antibody testing for celiac disease. Clin Chim Acta 2010; 411 (1–2) 13-17
  CrossrefPubMedGoogle Scholar
• 37 Kumar V, Bartholomew W, Shieh MT, Pierce C, Kaul N. Standardization of ELISA for the detection of anti-cardiolipin antibodies—effect of non-specific IgG binding. Immunol Invest 1991; 20 (7) 583-593
  CrossrefPubMedGoogle Scholar
• 38 Cowchock S, Fort J, Munoz S, Norberg R, Maddrey W. False positive ELISA tests for anticardiolipin antibodies in sera from patients with repeated abortion, rheumatologic disorders and primary biliary cirrhosis: correlation with elevated polyclonal IgM and implications for patients with repeated abortion. Clin Exp Immunol 1988; 73 (2) 289-294
  PubMedGoogle Scholar
• 39 Bizzaro N, Pasini P, Finco B. False-positive reactions for IgA anti-phospholipid and anti-beta(2)-glycoprotein I antibodies in patients with IgA monoclonal gammopathy. Clin Chem 1999; 45 (11) 2007-2010
  PubMedGoogle Scholar
• 40 Roubey RA. Antigenic specificities of antiphospholipid autoantibodies: implications for clinical laboratory testing and diagnosis of the antiphospholipid syndrome. Lupus 1996; 5 (5) 425-430
  PubMedGoogle Scholar
• 41 Smeenk RJ, Lucassen WA, Swaak TJ. Is anticardiolipin activity a cross-reaction of anti-DNA or a separate entity?. Arthritis Rheum 1987; 30 (6) 607-617
  CrossrefPubMedGoogle Scholar
• 42 Hunt JE, McNeil HP, Morgan GJ, Crameri RM, Krilis SA. A phospholipid-beta 2-glycoprotein I complex is an antigen for anticardiolipin antibodies occurring in autoimmune disease but not with infection. Lupus 1992; 1 (2) 75-81
  CrossrefPubMedGoogle Scholar
• 43 Pengo V, Biasiolo A, Fior MG. Autoimmune antiphospholipid antibodies are directed against a cryptic epitope expressed when beta 2-glycoprotein I is bound to a suitable surface. Thromb Haemost 1995; 73 (1) 29-34
  PubMedGoogle Scholar
• 44 Clinical and Laboratory Standards Institute (CLSI). Collection, Transport and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays: Approved Guideline-Fifth Edition. CLSI document H21–A5. Clinical and Laboratory Standards Institute: Wayne, PA; 2008
  Google Scholar
• 45 Pengo V, Tripodi A, Reber G , et al; Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardisation Committee of the International Society on Thrombosis and Haemostasis. Update of the guidelines for lupus anticoagulant detection. J Thromb Haemost 2009; 7 (10) 1737-1740
  CrossrefPubMedGoogle Scholar
• 46 Brandt JT, Triplett DA, Alving B, Scharrer I. Criteria for the diagnosis of lupus anticoagulants: an update. On behalf of the Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardisation Committee of the ISTH. Thromb Haemost 1995; 74 (4) 1185-1190
  Article in Thieme ConnectPubMedGoogle Scholar
• 47 Brien WF, Schaus MR, Cooper KE, O'Keefe BT, Inwood M. Lupus anticoagulant testing: effect of the platelet count on the activated partial thromboplastin time. Br J Biomed Sci 1993; 50 (2) 114-116
  PubMedGoogle Scholar
• 48 Tripodi A. Laboratory testing for lupus anticoagulants: a review of issues affecting results. Clin Chem 2007; 53 (9) 1629-1635
  CrossrefPubMedGoogle Scholar
• 49 Tripodi A, Biasiolo A, Chantarangkul V, Pengo V. Lupus anticoagulant (LA) testing: performance of clinical laboratories assessed by a national survey using lyophilized affinity-purified immunoglobulin with LA activity. Clin Chem 2003; 49 (10) 1608-1614
  CrossrefPubMedGoogle Scholar
• 50 Sangle NA, Rodgers GM, Smock KJ. Prevalence of heparin in samples submitted for lupus anticoagulant testing. Lab Hematol 2011; 17 (1) 6-11
  CrossrefPubMedGoogle Scholar
• 51 Jacobsen EM, Trettenes EJ, Wisløff F, Abildgaard U. Detection and quantification of lupus anticoagulants in plasma from heparin treated patients, using addition of polybrene. Thromb J 2006; 4: 3
  CrossrefPubMedGoogle Scholar
• 52 Gosselin RC, King JH, Janatpur KA, Dager WH, Larkin EC, Owings JT. Effects of pentasaccharide (fondaparinux) and direct thrombin inhibitors on coagulation testing. Arch Pathol Lab Med 2004; 128 (10) 1142-1145
  PubMedGoogle Scholar
• 53 Genzen JR, Miller JL. Presence of direct thrombin inhibitors can affect the results and interpretation of lupus anticoagulant testing. Am J Clin Pathol 2005; 124 (4) 586-593
  CrossrefPubMedGoogle Scholar
• 54 Favaloro EJ, Posen J, Ramakrishna R , et al. Factor V inhibitors: rare or not so uncommon? A multi-laboratory investigation. Blood Coagul Fibrinolysis 2004; 15 (8) 637-647
  CrossrefPubMedGoogle Scholar
• 55 Tripodi A, Mancuso ME, Chantarangkul V , et al. Lupus anticoagulants and their relationship with the inhibitors against coagulation factor VIII: considerations on the differentiation between the 2 circulating anticoagulants. Clin Chem 2005; 51 (10) 1883-1885
  CrossrefPubMedGoogle Scholar
• 56 Ballard HS, Nyamuswa G. Life-threatening haemorrhage in a patient with rheumatoid arthritis and a lupus anticoagulant coexisting with acquired autoantibodies against factor VIII. Br J Rheumatol 1993; 32 (6) 515-517
  CrossrefPubMedGoogle Scholar
• 57 Saxena R, Dhot PS, Saraya AK, Singh H, Malhotra OP. Simultaneous occurrence of factor VIIIC inhibitor and lupus anticoagulant. Am J Hematol 1993; 42 (2) 232-233
  CrossrefPubMedGoogle Scholar
• 58 Blanco AN, Alcira Peirano A, Grosso SH, Gennari LC, Pérez Bianco R, Lazzari MA. A chromogenic substrate method for detecting and titrating anti-factor VIII antibodies in the presence of lupus anticoagulant. Haematologica 2002; 87 (3) 271-278
  PubMedGoogle Scholar