γ′ fibrinogen: Evaluation of a new assay for study of associations with cardiovascular disease

Rehana S. Lovely, Steven C. Kazmierczak, Joseph M. Massaro, Ralph B. D'Agostino, Christopher J. O'Donnell, David H. Farrell

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Abstract

BACKGROUND: Studies of disease associations with γ′ fibrinogen, a newly emerging risk factor for cardiovascular disease, have been hampered by the lack of a standardized and well-characterized assay. METHODS: We developed an immunometric technique to measure γ′ fibrinogen concentrations in plasma and studied the clinical utility of this test in samples from healthy individuals enrolled in the Framingham Offspring Study and in a separate case/control study of coronary artery disease (CAD). Monoclonal antibody 2.G2.H9, specific for the unique carboxyl terminal peptide of the fibrinogen γ′ chain, was used as capture antibody. Sheep antihuman fibrinogen/horseradish peroxidase conjugate was used for detection, with 3,3′,5,5′-tetramethylbenzidine as substrate. We evaluated the linearity, imprecision, analytical specificity, and lower limit of quantification of the assay. We determined the reference interval for γ′ fibrinogen in healthy individuals from the Framingham Offspring Study (n = 2879) and quantified associations between γ′ fibrinogen and cardiovascular disease risk factors. The sensitivity and specificity of γ′ fibrinogen in evaluating CAD patients (n = 133) was determined with ROC curve analysis. RESULTS: The γ′ fibrinogen ELISA had within-run CVs of 13.4% at 0.127 g/L and 4.8% at 0.416 g/L. The limit of quantification at an imprecision of 20% was 0.10 g/L. The reference interval for healthy individuals was 0.088-0.551 g/L. ROC curve analysis of results from patients with CAD yielded an area under the curve of 0.76, with a diagnostic accuracy of 0.78 at a decision threshold of 0.30 g/L. CONCLUSIONS: γ′ Fibrinogen shows excellent utility for cardiovascular risk analysis.

Original languageEnglish
Pages (from-to)781-788
Number of pages8
JournalClinical Chemistry
Volume56
Issue number5
DOIs
StatePublished - 1 May 2010

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Fibrinogen
Assays
Cardiovascular Diseases
ROC Curve
Coronary Artery Disease
Risk analysis
Horseradish Peroxidase
Area Under Curve
Case-Control Studies
Sheep
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Plasmas
Sensitivity and Specificity
Peptides
Antibodies
Substrates

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Lovely, R. S., Kazmierczak, S. C., Massaro, J. M., D'Agostino, R. B., O'Donnell, C. J., & Farrell, D. H. (2010). γ′ fibrinogen: Evaluation of a new assay for study of associations with cardiovascular disease. Clinical Chemistry, 56(5), 781-788. https://doi.org/10.1373/clinchem.2009.138347
Lovely, Rehana S. ; Kazmierczak, Steven C. ; Massaro, Joseph M. ; D'Agostino, Ralph B. ; O'Donnell, Christopher J. ; Farrell, David H. / γ′ fibrinogen : Evaluation of a new assay for study of associations with cardiovascular disease. In: Clinical Chemistry. 2010 ; Vol. 56, No. 5. pp. 781-788.
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Lovely, RS, Kazmierczak, SC, Massaro, JM, D'Agostino, RB, O'Donnell, CJ & Farrell, DH 2010, 'γ′ fibrinogen: Evaluation of a new assay for study of associations with cardiovascular disease', Clinical Chemistry, vol. 56, no. 5, pp. 781-788. https://doi.org/10.1373/clinchem.2009.138347

γ′ fibrinogen : Evaluation of a new assay for study of associations with cardiovascular disease. / Lovely, Rehana S.; Kazmierczak, Steven C.; Massaro, Joseph M.; D'Agostino, Ralph B.; O'Donnell, Christopher J.; Farrell, David H.

In: Clinical Chemistry, Vol. 56, No. 5, 01.05.2010, p. 781-788.

Research output: Contribution to journalArticle

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T1 - γ′ fibrinogen

T2 - Evaluation of a new assay for study of associations with cardiovascular disease

AU - Lovely, Rehana S.

AU - Kazmierczak, Steven C.

AU - Massaro, Joseph M.

AU - D'Agostino, Ralph B.

AU - O'Donnell, Christopher J.

AU - Farrell, David H.

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N2 - BACKGROUND: Studies of disease associations with γ′ fibrinogen, a newly emerging risk factor for cardiovascular disease, have been hampered by the lack of a standardized and well-characterized assay. METHODS: We developed an immunometric technique to measure γ′ fibrinogen concentrations in plasma and studied the clinical utility of this test in samples from healthy individuals enrolled in the Framingham Offspring Study and in a separate case/control study of coronary artery disease (CAD). Monoclonal antibody 2.G2.H9, specific for the unique carboxyl terminal peptide of the fibrinogen γ′ chain, was used as capture antibody. Sheep antihuman fibrinogen/horseradish peroxidase conjugate was used for detection, with 3,3′,5,5′-tetramethylbenzidine as substrate. We evaluated the linearity, imprecision, analytical specificity, and lower limit of quantification of the assay. We determined the reference interval for γ′ fibrinogen in healthy individuals from the Framingham Offspring Study (n = 2879) and quantified associations between γ′ fibrinogen and cardiovascular disease risk factors. The sensitivity and specificity of γ′ fibrinogen in evaluating CAD patients (n = 133) was determined with ROC curve analysis. RESULTS: The γ′ fibrinogen ELISA had within-run CVs of 13.4% at 0.127 g/L and 4.8% at 0.416 g/L. The limit of quantification at an imprecision of 20% was 0.10 g/L. The reference interval for healthy individuals was 0.088-0.551 g/L. ROC curve analysis of results from patients with CAD yielded an area under the curve of 0.76, with a diagnostic accuracy of 0.78 at a decision threshold of 0.30 g/L. CONCLUSIONS: γ′ Fibrinogen shows excellent utility for cardiovascular risk analysis.

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