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Free light chain EQA: Analytical variability with clinical consequences - Data from 25 Italian EQA exercises
Fabio Pasotti1, Simona Da Molin1, Tiziana Bosoni2
1Regional Coordination Center for Laboratory Medicine of Lombardy Region, Milan, Italy.
Serum free light chain (FLC) assays show significant differences between methods, impacting monoclonal gammopathy diagnosis. Harmonized interpretation requires method-specific reference intervals and standardization pending international consensus.
Area of Science:
- Clinical Chemistry
- Immunology
- Biomarker Analysis
Background:
- Serum free light chains (FLC) κ and λ, and their ratio, are crucial biomarkers for monoclonal gammopathies.
- Current FLC assays exhibit significant inter-method variability, hindering standardized clinical interpretation.
Purpose of the Study:
- To evaluate the between-method variability of serum FLC assays using external quality assessment (EQA) data.
- To assess the impact of assay variability on the interpretation of FLC κ/λ ratios.
Main Methods:
- Analysis of five years (2021-2025) of Italian FLC EQA scheme data (2218 results).
- Application of ISO 13528 standards with robust statistics and method-specific peer groups.
- Comparison of Siemens N Latex and Binding Site Freelite assays using Bland-Altman analysis.
Main Results:
- Inter-laboratory precision within peer groups was acceptable (CV ≈ 10-20%).
- Marked between-method differences were observed: Freelite showed ~2x higher κ FLC (+94% bias) and lower λ FLC (-82% bias) compared to Siemens N Latex.
- Significant discrepancies in κ/λ ratio interpretation were identified, with biases persisting across concentration ranges.
Conclusions:
- Current FLC assays are not interchangeable, necessitating method-specific interpretation strategies.
- Universal reference intervals for κ/λ ratios are inappropriate due to assay variability.
- Method-specific cut-offs, consistent follow-up, and stratified EQA schemes are essential pending international standardization.
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