New combinational assay using soluble fibrin and d-dimer determinations: a promising strategy for identifying patients with suspected venous thromboembolism

Shahsoltan Mirshahi, Claudine Soria, Basile Kouchakji, Gérald Kierzek, Jeanne Yvonne Borg, Rémi Varin, Jean Chidiac, Ludovic Drouet, Massoud Mirshahi, Jeannette Soria, Shahsoltan Mirshahi, Claudine Soria, Basile Kouchakji, Gérald Kierzek, Jeanne Yvonne Borg, Rémi Varin, Jean Chidiac, Ludovic Drouet, Massoud Mirshahi, Jeannette Soria

Abstract

Aim: To establish a new and reliable assay for quantification of the soluble fibrin (SF) in combination with that of D-dimer for early diagnosis of venous thromboembolism.

Methods and samples: The SF assay is based on D-dimer generated after incubation of plasma with tissue-type plasminogen activator (t-PA). SF and standard D-dimer assays, run in blind, were used to test 119 untreated outpatients with clinically suspected deep-vein thrombosis (DVT, 49 patients) or pulmonary embolism (PE, 70 patients) consulting at the emergency unit of the hospital. Thromboses were confirmed by current imaging methods such as ultrasonography, scintigraphy, computed tomographic pulmonary angiography (CTPA) and ventilation/perfusion scan.

Results: SF assay was validated in 270 healthy volunteers [51.8% males; mean age years ± SD: 41±13; age range 19 to 65]. Among these normal plasmas, SF levels were ≤200 ng/mL in 97.8% of them, and 200-250 ng/mL in the remainder [26-46 years old; 50% males]. ROC curves were used to determine the SF cut-off value for plasma SF positivity, which was found to be 300 ng/mL. In patients with suspected venous thromboembolism, SF sensitivities for DVT and PE (92% and 94%, respectively) were comparable to those of D-dimer (96% and 94%), whereas SF specificities (86% and 95%) were higher than those of D-dimer (50% and 54%). Positive-predictive values for SF (89% and 94%) were again higher than those of D-dimer (70% and 65%) in DVT and PE. The amount of circulating SF normalized rapidly after anticoagulant therapy.

Conclusion: Results from this small group of patients suggest that the evaluation of plasma SF, in combination with that of D-dimer, represents a potentially useful tool for the early diagnosis of venous thromboembolism, provided that the patients have not been treated previously by anticoagulants.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Evolution of SF and D-dimer…
Figure 1. Evolution of SF and D-dimer levels in patients under anticoagulant therapy.
SF and D-dimer (D-Di) concentrations were determined daily in patients before and after starting anticoagulant therapy.
Figure 2. ROC curves for DVT and…
Figure 2. ROC curves for DVT and PE.
ROC Curves established for Pulmonary Embolism and for Deep Vein Thrombosis Open squares : Sensitivity Closed square : Specificity.
Figure 3. Distribution of the levels of…
Figure 3. Distribution of the levels of SF and D-dimer in patients with suspected pulmonary embolism (PE) or deep vein thrombosis.
Evaluation of median and 25th and 75th percentiles. (A) Soluble fibrin concentration. (B) D-dimer concentration. •/Closed circles: patients with PE or DVT; ○/open circles, patients without PE or DVT. The bars depict the median with interquartile ranges. The double broken bars represent the upper normal value.

References

    1. Goldhaber SZ (1992) Pulmonary embolism thrombolysis: a clarion call for international collaboration. J Am Coll Cardiol 9: 246–7.
    1. Rosendaal FR (1999) Venous thrombosis: a multicausal disease. Lancet 353: 1167–73.
    1. Bounameaux H, Khabiri E, Huber O, Schneider PA, Didier D, et al. (1992) Value of liquid crystal contact thermography and plasma level of D-dimer for screening of deep venous thrombosis following general abdominal surgery. Thromb Haemost 67: 603–6.
    1. Duet M, Benelhadj S, Kedra W, Vilain D, Ajzenberg C, et al. (1998) A new quantitative D-dimer assay appropriate in emergency: reliability of the assay for pulmonary embolism exclusion diagnosis. Thromb Res 91: 1–5.
    1. Bounameaux H, Cirafici P, de Moerloose P, Schneider PA, Slosman D, et al. (1991) Measurement of D-dimer in plasma as diagnostic aid in suspected pulmonary embolism. Lancet 337: 196–200.
    1. Bounameaux H, Schneider PA, Slosman D, de Moerloose P, Reber G (1990) Plasma D-dimer in suspected pulmonary embolism: a comparison with pulmonary angiography and ventilation–perfusion scintigraphy. Blood Coagul Fibrinolysis 1: 1577–9.
    1. Lippi G, Franchini M, Biasiutti C, Dellagiacoma G, Salvagno GL, et al. (2007) Increased D-dimer value and occult cancer in the absence of detectable thrombosis. Haematologica 92: 53–5.
    1. Biancone L, Scopinaro F, Maletta M, Monteleone G, Luzza F, Banci M, et al. (1994) Circulating D dimer in inflammatory bowel disease. Ital J Gastroenterol 26: 116–20.
    1. Ginsberg JS, Siragusa S, Douketis J, Johnston M, Moffat K, et al. (1996) Evaluation of a soluble fibrin assay in patients with suspected pulmonary embolism. Thromb Haemost 75: 551–4.
    1. Hetland O, Knudsen A, Dickstein K, Nilsen DW (2002) Characteristics and prognostic impact of plasma fibrin monomer (soluble fibrin) in patients with coronary artery disease. Blood Coagul Fibrinolysis 13: 301–8.
    1. Rötker J, Preissner KT, Müller-Berghaus G (1986) Soluble fibrin consists of fibrin oligomers of heterogeneous distribution. Eur J Biochem. 155: 583–8.
    1. Brummel KE, Butenas S, Mann KG (1999) An integrated study of fibrinogen during blood coagulation. J Biol Chem 274: 22862–70.
    1. Eisenberg PR, Lucore C, Kaufman L, Sobel BE, Jaffe AS, et al. (1990) Fibrinopeptide A levels indicative of pulmonary vascular thrombosis in patients with primary pulmonary hypertension. Circulation 82: 841–7.
    1. LaCapra S, Arkel YS, Ku DH, Gibson D, Lake C, et al. (2000) The use of thrombus precursor protein, D-dimer, prothrombin fragment 1.2, and thrombin antithrombin in the exclusion of proximal deep vein thrombosis and pulmonary embolism. Blood Coagul Fibrinolysis 11: 371–7.
    1. Mac Gillavry MR, Sanson BJ, de Monye W, Lijmer JG, Huisman MV, et al. (2000) Use of a new monoclonal antibody-based enzyme immunoassay for soluble fibrin to exclude pulmonary embolism. ANTELOPE-Study Group. Thromb Haemost 84: 474–7.
    1. Iversen LH, Thorlacius-Ussing O (1996) Markers of fibrin formation in colorectal surgery: fibrinopeptide A and soluble fibrin. Thromb Res (81) 569–576.
    1. Lehman CM, Wilson LW, Rodgers GM (2004) Analytic Validation and Clinical evaluation of the STA LIATEST Immunoturbidimetric D-Dimer Assay for the Diagnosis of Disseminated Intravascular Coagulation. Am J Clin Pathol 122: 178–184.
    1. Mosesson MW, Finlayson JS (1963) Subfractions of human fibrinogen: preparation and analysis. J Lab Clin Med 62: 663–674.
    1. Hanley JA, McNeil BJ (1982) The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology 43: 29–36.
    1. Hanley JA McNeil BJ (1983) A method of comparing the area under two ROC curves derived from the same cases. Radiology 148: 839–843.
    1. Flores J, García-Avello A, Flores VM, Navarro JL, Canseco F, et al. (2003) Tissue Plasminogen Activator Plasma Levels as a Potential Diagnostic Aid in Acute Pulmonary Embolism. Arch Pathol Lab Med 127: 310–315.
    1. Chandler WL, Trimble SL, Loo SC, Mornin D (1990) Effect of PAI-1 levels on the molar concentrations of active tissue plasminogen activator (t-PA) and t-PA/PAI-1 complex in plasma. Blood 76: 930–7.
    1. Crowther MA, Roberts J, Roberts R, Johnston M, Stevens P, et al. (2001) Fibrinolytic variables in patients with recurrent venous thrombosis: a prospective cohort study. Thromb Haemost 85: 390–394.
    1. Mirshahi SS, Pujade-Lauraine E, Soria C, Mirshahi M, Fretault J, et al. (1992) D-dimer and CA 125 levels in patients with ovarian cancer during antineoplastic therapy. Prognostic significance for the success of anti-cancer treatment. Cancer 69: 2289–92.
    1. Dirix LY, Salgado R, Weytjens R, Colpaert C, Benoy I, et al. (2002) Plasma fibrin D-dimer levels correlate with tumour volume, progression rate and survival in patients with metastatic breast cancer. Br J Cancer 86: 389–95.
    1. Bounameaux H, Schneider PA, Slosman D, De Moerloose P, Reber G (1991) Value of the plasma measurement of D-dimer in the diagnosis of venous thromboembolism. J Mal Vasc 16: 133–6.
    1. Wells PS, Anderson DR, Rodger M, Stiell I, Dreyer JF, et al. (2001) Excluding pulmonary embolism at the bedside without diagnostic imaging: management of patients with suspected pulmonary embolism presenting to the emergency department by using a simple clinical model and D-dimer. Ann Intern Med 135: 98–107.
    1. Hogg PJ, Jackson CM (1989) Fibrin monomer protects thrombin from inactivation by heparin-antithrombin III: implications for heparin efficacy. Proc Natl Acad Sci U S A 86: 3619–3623.
    1. Soria J, Soria C, Caen JP (1983) A new type of congenital dysfibrinogenaemia with defective fibrin lysis—Dusard syndrome: possible relation to thrombosis. Br J Haematol 53: 575–86.
    1. Dempfle CE, Wurst M, Smolinski M, Lorenz S, Osika A, et al. (2004) Use of soluble fibrin antigen instead of D-dimer as fibrin-related marker may enhance the prognostic power of the ISTH overt DIC score. Thromb Haemost 91: 812–8.
    1. Derhaschnig U, Laggner AN, Roggla M, Hirschl MM, Kapiotis S, et al. (2002) Evaluation of coagulation markers for early diagnosis of acute coronary syndromes in the emergency room. Clin Chem 48: 1924–30.
    1. Arkel YS, Ku DH, Le P, Carr AM (2001) Comparison of a test for soluble fibrin polymer (TpP) with a standard quantitative ELISA for D-dimer in patients, without current thrombosis, who have cancer or renal disease. Thromb Haemost 86: 1127–8.
    1. Arkel YS, Paidas MJ, Ku DH (2002) The use of coagulation activation markers (soluble fibrin polymer, TpP, prothrombin fragment 1.2, thrombin-antithrombin, and D-dimer) in the assessment of hypercoagulability in patients with inherited and acquired prothrombotic disorders. Blood Coagul Fibrinolysis 13: 199–205.
    1. Brimble KS, Ginsberg JS (1997) Evaluation of the combination of a bedside D-dimer assay and enzyme-linked immunosorbent soluble fibrin assay in patients with suspected venous thromboembolism. Thromb Res 88: 291–7.
    1. Dempfle CE, Dollman M, Lill H, Puzzovio D, Dessauer A, et al. (1993) Binding of a new monoclonal antibody against N-terminal heptapeptide of fibrin alpha-chain to fibrin polymerization site 'A': effects of fibrinogen and fibrinogen derivatives, and pretreatment of samples with NaSCN. Fibrinolysis 4: 79–86.
    1. Dempfle CE, Pfitzner SA, Dollman M, Huck K, Stehle G, et al. (1995) Comparison of immunological and functional assays for measurement of soluble fibrin. Thromb Haemost 74: 673–9.
    1. Dempfle CE, Zips S, Ergul H, Heene DL (2001) FACT Study Group (2001) The fibrin assay comparison trial (FACT): correlation of soluble fibrin assays with D-dimer. Thromb Haemost 86: 1204–9.
    1. Ginsberg JS, Siragusa S, Douketis J, Johnston M, Moffat K, et al. (1995) Evaluation of a soluble fibrin assay in patients with suspected deep vein thrombosis. Thromb Haemost 74: 833–6.
    1. Ginsberg JS, Siragusa S, Douketis J, Johnston M, Moffat K, et al. (1996) Evaluation of a soluble fibrin assay in patients with suspected pulmonary embolism. Thromb Haemost 75: 551–4.
    1. Hetland O, Knudsen A, Dickstein K, Nilsen DW (2002) Characteristics and prognostic impact of plasma fibrin monomer (soluble fibrin) in patients with coronary artery disease. Blood Coagul Fibrinolysis 13: 301–8.
    1. Koga S (2004) A novel molecular marker for thrombus formation and life prognosis – clinical usefulness of measurement of soluble fibrin monomer-fibrinogen complex (SF). Rinsho Byori 52: 355–61.
    1. Nakahara K, Kazahaya Y, Shintani Y, Yamazumi K, Eguchi Y, et al. (2003) Measurement of soluble fibrin monomer-fibrinogen complex in plasmas derived from patients with various underlying clinical situations. Thromb Haemost 89: 832–6.
    1. Nieuwenhuizen W, Hoegee-De Nobel E, Laterveer R (1992) A rapid monoclonal antibody-based enzyme immunoassay (EIA) for the quantitative determination of soluble fibrin in plasma. Thromb Haemost 68: 273–7.
    1. Nieuwenhuizen W (1993) Soluble fibrin as a molecular marker for a pre-thrombotic state: a mini-review. Blood Coagul Fibrinolysis 4: 93–6.
    1. Okajima K, Uchiba M, Murakami K, Okabe H, Takatsuki K (1996) Determination of plasma soluble fibrin using a new ELISA method in patients with disseminated intravascular coagulation. Am J Hematol 51: 186–91.
    1. Reber G, Bounameaux H, Perrier A, de Moerloose P (1999) Performances of the fibrin monomer test for the exclusion of pulmonary embolism in symptomatic outpatients. Thromb Haemost 81: 221–3.
    1. Scarano L, Prandoni P, Gavasso S, Gomiero W, Carraro G, et al. (1999) Failure of soluble fibrin polymers in the diagnosis of clinically suspected deep venous thrombosis. Blood Coagul Fibrinolysis 10: 245–50.
    1. Wada H, Sase T, Matsumoto T, Kushiya F, Sakakura M, et al. (2003) Increased soluble fibrin in plasma of patients with disseminated intravascular coagulation. Clin Appl Thromb Hemost 9: 233–40.

Source: PubMed

Sponsor e collaboratori

Condizioni mediche

Interventi farmacologici

3
Sottoscrivi