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Related Experiment Videos

Variability in plasma prothrombin concentration: implications for use in epidemiology

A M Smiles1, E M Macy, P A Sakkinen

  • 1Department of Pathology, University of Vermont, Colchester 05446, USA.

Blood Coagulation & Fibrinolysis : an International Journal in Haemostasis and Thrombosis
|November 18, 1998
PubMed
Summary

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A new enzyme-linked immunosorbent assay (ELISA) accurately measures prothrombin levels, a key thrombosis risk factor. This cost-effective method is suitable for large-scale population studies, overcoming limitations of traditional clotting assays.

Area of Science:

  • Biochemistry
  • Clinical Chemistry
  • Hematology

Background:

  • Prothrombin is increasingly recognized as a significant risk factor for thrombosis.
  • Current methods for measuring prothrombin levels, such as plasma-based clotting assays (factor IIc), are costly and complex for population research.

Purpose of the Study:

  • To develop and validate a novel enzyme-linked immunosorbent assay (ELISA) for quantifying prothrombin levels.
  • To assess the assay's suitability for population-based research by evaluating its precision, accuracy, and biovariability.

Main Methods:

  • Development of an ELISA using purified prothrombin antigen and polyclonal anti-prethrombin-1 IgG.
  • Validation included assessing coefficients of variation (CV) in quality control plasmas, analytical recovery, and correlation with factor IIc assay results.

Related Experiment Videos

  • Biovariability studies determined analytical variability (CV(A)), within-subject variability (CV(I)), and between-subject variability (CV(G)).

    • Main Results:

    • The developed ELISA demonstrated high precision with CVs below 6.5% in quality control plasmas and an average analytical recovery of 103.8%.
    • ELISA results showed good correlation with factor IIc assay (r=0.75).
    • Biovariability analysis indicated an index of individuality of 0.50, suggesting the assay is well-suited for population studies. No significant cross-reactivity or diurnal variation was observed.

    Conclusions:

    • The developed ELISA provides a reliable, cost-effective, and precise method for measuring prothrombin levels.
    • The assay's performance characteristics, particularly its low variability and suitability for population studies, address limitations of existing methods.
    • This ELISA facilitates large-scale research into the role of prothrombin in thrombosis risk.