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A soluble fibrin standard: comparable dose-response with immunologic and functional assays
B I McCarron1, V J Marder, J J Kanouse
1Vascular Medicine Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, New York, USA.
Thrombosis and Haemostasis
|August 24, 1999
Summary
A new soluble fibrin (SF) standard has been developed for laboratory assays. This preparation effectively normalizes results across different commercial assays, improving diagnostic accuracy for soluble fibrin levels.
Area of Science:
- Biochemistry
- Hematology
- Clinical Chemistry
Background:
- Accurate measurement of soluble fibrin (SF) is crucial for diagnosing and monitoring various hemostatic disorders.
- Current diagnostic assays for SF often lack standardization, leading to variability in results.
- A reliable standard is needed to ensure consistency and comparability across different laboratory tests.
Purpose of the Study:
- To develop and characterize a soluble fibrin (SF) preparation suitable for use as a standard in laboratory assays.
- To evaluate the performance of this SF standard across multiple commercially available diagnostic kits.
- To assess the potential of the SF standard to normalize results from disparate internal calibrators used in different assays.
Main Methods:
- Preparation of the SF standard involved reacting factor XIII-free fibrinogen with thrombin, followed by hirudin neutralization and solubilization in acetic acid.
- Characterization of the SF preparation using SDS-PAGE confirmed the expected polypeptide structure, including loss of fibrinopeptides and absence of crosslinking.
- The SF standard was tested in pooled normal plasma across a range of concentrations (12.5–340 µg/ml) using four different commercial assays (ELISA, latex agglutination, t-PA cofactor activity).
Main Results:
- The SF standard demonstrated distinct dose-response relationships when tested with the four evaluated assays.
- Log-transformation of assay results generated parallel lines, indicating consistent performance of the SF standard across different assay methodologies.
- Calculated absolute SF concentrations varied based on manufacturers' calibrators, highlighting the need for a universal standard.
Conclusions:
- The developed soluble fibrin preparation is suitable for use as a standard in laboratory assays evaluating SF levels.
- This SF standard can effectively normalize the results obtained from different commercial assays, overcoming the limitations of proprietary internal calibrators.
- Implementation of this SF standard is expected to improve the reliability and comparability of SF measurements in clinical diagnostics.