Validation, Calibration, and Specificity of Quantitative D-Dimer Assays

ABSTRACT

Assays for D-dimer antigen are based on monoclonal antibodies reactive with epitopes found on fibrin fragment D-dimer but not on fibrinogen fragment D, other fibrinogen degradation products, or native fibrinogen. The antibodies react with conformational epitopes generated by factor XIII-induced linkage of the C-terminal appendages of the fibrin γ-chains of adjacent D-domains within a fibrin polymer. For some monoclonal antibodies, degradation of the cross-linked fibrin compound by plasmin is an additional requirement for the generation of the epitope. In clinical plasma samples, D-dimer antigen assays detect an array of fibrin compounds of different molecular weights, including fibrin fragment D-dimer as well as higher-molecular-weight fibrin degradation products and fibrin X-oligomers. Most D-dimer antigen represents cross-linked soluble fibrin present in circulation rather than degradation products from particulate clots. Due to differences in epitope reactivity, harmonization of D-dimer antigen assays can only be achieved with standard preparations containing a similar variety of cross-linked fibrin compounds. Assay technologies include manual latex agglutination assays, automated latex-enhanced light-scattering immunoassays, enzyme-linked immunoassays, and others.

D-dimer - fibrinogen/fibrin degradation products - fibrinogen - thrombin - factor XIII

REFERENCES

• 1 Hillyard C J, Blake A S, Wilson K et al.. A latex agglutination assay for D dimer: evaluation and application to the diagnosis of thrombotic disease.  Clin Chem. 1987;  33 1837-1840
• 2 Rowbotham B J, Carroll P, Whitaker A N et al.. Measurement of crosslinked fibrin derivatives-use in the diagnosis of venous thrombosis.  Thromb Haemost. 1987;  57 59-61
• 3 Goldhaber S Z, Vaughan D E, Tumeh S S, Loscalzo J. Utility of cross-linked fibrin degradation products in the diagnosis of pulmonary embolism.  Am Heart J. 1988;  116 505-508
• 4 Bounameaux H, Slosman D, de Moerloose P, Reber G. Diagnostic value of plasma D-dimer in suspected pulmonary embolism.  Lancet. 1988;  2 628-629
• 5 Lane D A, Preston F E, VanRoss M E, Kakkar V V. Characterization of serum fibrinogen and fibrin fragments produced during disseminated intravascular coagulation.  Br J Haematol. 1978;  40 609-615
• 6 Kobayashi T, Terao T. Preeclampsia as chronic disseminated intravascular coagulation. Study of two parameters: thrombin-antithrombin III complex and D-dimers.  Gynecol Obstet Invest. 1987;  24 170-178
• 7 Ballegeer V, Mombaerts P, Declerck P J, Spitz B, Van Assche F A, Collen D. Fibrinolytic response to venous occlusion and fibrin fragment D-dimer levels in normal and complicated pregnancy.  Thromb Haemost. 1987;  58 1030-1032
• 8 Trofatter Jr K F, Howell M L, Greenberg C S, Hage M L. Use of the fibrin D-dimer in screening for coagulation abnormalities in preeclampsia.  Obstet Gynecol. 1989;  73 435-440
• 9 Khoo S K, Rylatt D B, Parsons P et al.. Serial D-dimer levels in the assessment of tumor mass and clinical outcome in ovarian cancer.  Gynecol Oncol. 1988;  29 188-198
• 10 Olexa S A, Budzynski A Z. Evidence for four different polymerization sites involved in human fibrin formation.  Proc Natl Acad Sci USA. 1980;  77 1374-1378
• 11 Weisel J W, Veklich Y, Gorkun O. The sequence of cleavage of fibrinopeptides from fibrinogen is important for protofibril formation and enhancement of lateral aggregation in fibrin clots.  J Mol Biol. 1993;  232 285-297
• 12 Lounes K C, Ping L, Gorkun O V, Lord S T. Analysis of engineered fibrinogen variants suggests that an additional site mediates platelet aggregation and that “B-b” interactions have a role in protofibril formation.  Biochemistry. 2002;  41 5291-5299
• 13 Greenberg C S, Achyuthan K E, Rajagopalan S, Pizzo S V. Characterization of the fibrin polymer structure that accelerates thrombin cleavage of plasma factor XIII.  Arch Biochem Biophys. 1988;  262 142-148
• 14 Yee V C, Pedersen L C, Bishop P D, Stenkamp R E, Teller D C. Structural evidence that the activation peptide is not released upon thrombin cleavage of factor XIII.  Thromb Res. 1995;  78 389-397
• 15 Muszbek L, Yee V C, Hevessy Z. Blood coagulation factor XIII: structure and function.  Thromb Res. 1999;  94 271-305
• 16 Brummel K E, Butenas S, Mann K G. An integrated study of fibrinogen during blood coagulation.  J Biol Chem. 1999;  274 22862-22870
• 17 Siebenlist K R, Meh D A, Mosesson M W. Protransglutaminase (factor XIII) mediated crosslinking of fibrinogen and fibrin.  Thromb Haemost. 2001;  86 1221-1228
• 18 Dempfle C E, Argiriou S, Alesci S et al.. Fibrin formation and proteolysis during ancrod treatment. Evidence for des-A-profibrin formation and thrombin independent factor XIII activity.  Ann N Y Acad Sci. 2001;  936 210-214
• 19 Dempfle C E, Argiriou S, Kucher K, Muller-Peltzer H, Rubsamen K, Heene D L. Analysis of fibrin formation and proteolysis during intravenous administration of ancrod.  Blood. 2000;  96 2793-2802
• 20 Spraggon G, Everse S J, Doolittle R F. Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin.  Nature. 1997;  389 455-462
• 21 Veklich Y, Ang E K, Lorand L, Weisel J W. The complementary aggregation sites of fibrin investigated through examination of polymers of fibrinogen with fragment E.  Proc Natl Acad Sci USA. 1998;  95 1438-1442
• 22 Mosesson M W, Siebenlist K R, Hernandez I, Wall J S, Hainfeld J F. Fibrinogen assembly and crosslinking on a fibrin fragment E template.  Thromb Haemost. 2002;  87 651-658
• 23 Siebenlist K R, Meh D A, Mosesson M W. Position of gamma-chain carboxy-terminal regions in fibrinogen/fibrin cross-linking mixtures.  Biochemistry. 2000;  39 14171-14175
• 24 Mosesson M W, Siebenlist K R, Meh D A, Wall J S, Hainfeld J F. The location of the carboxy-terminal region of gamma chains in fibrinogen and fibrin D domains.  Proc Natl Acad Sci USA. 1998;  95 10511-10516
• 25 Mosesson M W, Siebenlist K R, Meh D A. The structure and biological features of fibrinogen and fibrin.  Ann N Y Acad Sci. 2001;  936 11-30
• 26 Siebenlist K R, Mosesson M W. Factors affecting gamma-chain multimer formation in cross-linked fibrin.  Biochemistry. 1992;  31 936-941
• 27 Siebenlist K R, Mosesson M W. Progressive cross-linking of fibrin gamma chains increases resistance to fibrinolysis.  J Biol Chem. 1994;  269 28414-28419
• 28 Wiman B, Mellbring G, Ranby M. Plasminogen activator release during venous stasis and exercise as determined by a new specific assay.  Clin Chim Acta. 1983;  127 279-288
• 29 Mosesson M W, Siebenlist K R, Voskuilen M, Nieuwenhuizen W. Evaluation of the factors contributing to fibrin-dependent plasminogen activation.  Thromb Haemost. 1998;  79 796-801
• 30 Mosesson M W, Siebenlist K R, Voskuilen M, Nieuwenhuizen W. The effect of fibrinogen polymerization and crosslinking on exposure of fibrin-specific epitopes and t-PA mediated plasminogen activation.  Ukr Biokhim Zh. 1996;  68 37-38
• 31 Halvorsen S, Skjonsberg O H, Godal H C. The stimulatory effect of soluble fibrin on plasminogen activation by tissue plasminogen activator as studied by the Coa-set Fibrin Monomer test.  Thromb Res. 1991;  61 453-461
• 32 Halvorsen S, Skjonsberg O H, Godal H C. The stimulatory capacity of soluble fibrin prepared from high and low molecular weight fibrinogen on plasminogen activation.  Blood Coagul Fibrinolysis. 1993;  4 133-137
• 33 Dempfle C E, Alesci S, Kucher K, Muller-Peltzer H, Rubsamen K, Borggrefe M. Plasminogen activation without changes in tPA and PAI-1 in response to subcutaneous administration of ancrod.  Thromb Res. 2001;  104 433-438
• 34 Gaffney P J. Distinction between fibrinogen and fibrin degradation products in plasma.  Clin Chim Acta. 1975;  65 109-115
• 35 Francis C W, Marder V J, Barlow G H. Plasmic degradation of crosslinked fibrin. Characterization of new macromolecular soluble complexes and a model of their structure.  J Clin Invest. 1980;  66 1033-1043
• 36 Gaffney P J, Lane D A, Kakkar V V, Brasher M. Characterisation of a soluble D dimer-E complex in crosslinked fibrin digests.  Thromb Res. 1975;  7 89-99
• 37 Olexa S A, Budzynski A Z. Primary soluble plasmic degradation product of human cross-linked fibrin. Isolation and stoichiometry of the (DD)E complex.  Biochemistry. 1979;  18 991-995
• 38 Marder V J, Budzynski A Z, Barlow G H. Comparison of the physicochemical properties of fragment D derivatives of fibrinogen and fragment D-D of cross-linked fibrin.  Biochim Biophys Acta. 1976;  427 1-14
• 39 Walker J B, Nesheim M E. The molecular weights, mass distribution, chain composition, and structure of soluble fibrin degradation products released from a fibrin clot perfused with plasmin.  J Biol Chem. 1999;  274 5201-5212
• 40 Veklich Y, Francis C W, White J, Weisel J W. Structural studies of fibrinolysis by electron microscopy.  Blood. 1998;  92 4721-4729
• 41 Kornberg A, Francis C W, Marder V J. Plasma crosslinked fibrin polymers: quantitation based on tissue plasminogen activator conversion to D-dimer and measurement in normal and patients with acute thrombotic disorders.  Blood. 1992;  80 709-717
• 42 Cierniewski C S, Janiak A, Nowak P, Augustyniak W. Reactivity of fibrinogen derivatives with antisera to human fibrin D-dimer and its gamma-gamma chain remnant.  Thromb Haemost. 1982;  48 33-37
• 43 Matsushima A, Takahama Y, Inada Y. Preparation of guinea pig antiserum to quantitate serum fragment D-dimer derived from crosslinked fibrin.  Thromb Res. 1982;  27 111-115
• 44 Rylatt D B, Blake A S, Cottis L E et al.. An immunoassay for human D dimer using monoclonal antibodies.  Thromb Res. 1983;  31 767-778
• 45 Devine D V, Greenberg C S. Monoclonal antibody to fibrin D-dimer (DD-3B6) recognizes an epitope on the gamma-chain of fragment D.  Am J Clin Pathol. 1988;  89 663-666
• 46 Wylie F G, Walsh T P. Partial identification of the epitope on D-dimer for monoclonal antibody, DD-3B6/22.  Blood Coagul Fibrinolysis. 1997;  8 87-96
• 47 Gaffney P J, Edgell T, Creighton-Kempsford L J, Wheeler S, Tarelli E. Fibrin degradation product (FnDP) assays: analysis of standardization issues and target antigens in plasma.  Br J Haematol. 1995;  90 187-194
• 48 Pfitzner S A, Dempfle C E, Matsuda M, Heene D L. Fibrin detected in plasma of patients with disseminated intravascular coagulation by fibrin-specific antibodies consists primarily of high molecular weight factor XIIIa-crosslinked and plasmin-modified complexes partially containing fibrinopeptide A.  Thromb Haemost. 1997;  78 1069-1078
• 49 Gron B, Bennick A, Nieuwenhuizen W, Brosstad F. Normal and fibrinaemic patient plasma contain high-molecular weight crosslinked fibrin(ogen) derivatives with intact fibrinopeptide A.  Thromb Res. 1990;  57 259-270
• 50 Francis C W, Marder V J. Degradation of cross-linked fibrin by human leukocyte proteases.  J Lab Clin Med. 1986;  107 342-352
• 51 Matsuda M, Terukina S, Yamazumi K, Maekawa H, Soe G. A monoclonal antibody that recognizes the NH2-terminal conformation of fragment D.  Excerpta Medica. 1990;  892 43-48
• 52 Brenner B, Francis C W, Marder V J. The role of soluble cross-linked fibrin in D dimer immunoreactivity of plasmic digests.  J Lab Clin Med. 1989;  113 682-688
• 53 Dempfle C E, Zips S, Ergul H, Heene D L. The Fibrin Assay Comparison Trial (FACT): evaluation of 23 quantitative D-dimer assays as basis for the development of D-dimer calibrators. FACT study group.  Thromb Haemost. 2001;  85 671-678
• 54 Francis C W, Doughney K, Brenner B, Klingbiel K, Marder V J. Increased immunoreactivity of plasma after fibrinolytic activation in an anti-DD ELISA system. Role of soluble crosslinked fibrin polymers.  Circulation. 1989;  79 666-673
• 55 Brenner B, Francis C W, Totterman S et al.. Quantitation of venous clot lysis with the D-dimer immunoassay during fibrinolytic therapy requires correction for soluble fibrin degradation.  Circulation. 1990;  81 1818-1825
• 56 Bounameaux H, Schneider P A, Reber G, de Moerloose P, Krahenbuhl B. Measurement of plasma D-dimer for diagnosis of deep venous thrombosis.  Am J Clin Pathol. 1989;  91 82-85
• 57 Elias A, Aptel I, Huc B et al.. D-dimer test and diagnosis of deep vein thrombosis: a comparative study of 7 assays.  Thromb Haemost. 1996;  76 518-522
• 58 Froehling D A, Elkin P L, Swensen S J, Heit J A, Pankratz V S, Ryu J H. Sensitivity and specificity of the semiquantitative latex agglutination D-dimer assay for the diagnosis of acute pulmonary embolism as defined by computed tomographic angiography.  Mayo Clin Proc. 2004;  79 164-168
• 59 Pittet J L, de Moerloose P, Reber G et al.. VIDAS D-dimer: fast quantitative ELISA for measuring D-dimer in plasma.  Clin Chem. 1996;  42 410-415
• 60 Vissac A M, Grimaux M, Chartier S, Chan F A, Chambrette B, Amiral J. A new sensitive membrane based ELISA technique for instantaneous D. Dimer evaluation in emergency.  Thromb Res. 1995;  78 341-352
• 61 Dale S, Gogstad G O, Brosstad F et al.. Comparison of three D-dimer assays for the diagnosis of DVT:ELISA, latex and an immunofiltration assay (NycoCard D-Dimer).  Thromb Haemost. 1994;  71 270-274
• 62 Bucek R A, Quehenberger P, Feliks I, Handler S, Reiter M, Minar E. Results of a new rapid D-dimer assay (cardiac D-dimer) in the diagnosis of deep vein thrombosis.  Thromb Res. 2001;  103 17-23
• 63 Dempfle C, Schraml M, Besenthal I et al.. Multicentre evaluation of a new point-of-care test for the quantitative determination of D-dimer.  Clin Chim Acta. 2001;  307 211-218
• 64 John M A, Elms M J, O'Reilly E J, Rylatt D B, Bundesen P G, Hillyard C J. The simpliRED D dimer test: a novel assay for the detection of crosslinked fibrin degradation products in whole blood.  Thromb Res. 1990;  58 273-281
• 65 Adema E, Gebert U. Pooled patient samples as reference material for D-Dimer.  Thromb Res. 1995;  80 85-88
• 66 Nieuwenhuizen W. A reference material for harmonisation of D-dimer assays. Fibrinogen Subcommittee of the Scientific and Standardization Committee of the International Society of Thrombosis and Haemostasis.  Thromb Haemost. 1997;  77 1031-1033