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Interlaboratory comparison of current high-performance methods for HbA2
R Paleari1, B Gulbis, F Cotton
1CIRME, Dip. di Scienze e Tecnologie Biomediche, Università degli Studi di Milano, Italy.
International Journal of Laboratory Hematology
|February 11, 2012
Summary
Routine methods for measuring hemoglobin A2 (HbA2) show poor alignment, with significant variability in results. Better standardization of HbA2 measurement procedures is needed for accurate clinical interpretation.
Area of Science:
- Clinical Chemistry
- Hematology
- Analytical Chemistry
Background:
- External quality surveys reveal significant variability in hemoglobin A2 (HbA2) quantitation.
- Limited data exist on the alignment between different HbA2 measurement methods.
Purpose of the Study:
- To compare the performance of leading High-Performance Liquid Chromatography (HPLC) and Capillary Electrophoresis (CE) methods for HbA2 determination.
- To assess the alignment and potential interferences of various HbA2 quantitation techniques.
Main Methods:
- Analyzed 80 blood samples from normal subjects and beta-thalassemia carriers using multiple HPLC and CE systems.
- Included patient samples with clinically relevant hemoglobin variants (HbC, HbD, HbE, HbS, δ-chain variants) for interference testing.
Main Results:
- Within-run imprecision for HbA2 measurement ranged from 0.5% to 4.4% (HPLC) and 1.2% to 4.4% (CE).
- Methods showed high correlation (r=0.974-0.997) but exhibited biases.
- Both HPLC and CE methods demonstrated variable interference from abnormal hemoglobins.
Conclusions:
- A significant lack of alignment was observed among routine HbA2 measurement methods.
- There is a clear need for improved standardization in HbA2 measurement procedures to ensure reliable results.
